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African Journal of Food Science

June 2018 ISSN 1996-0794 DOI: 10.5897/AJFS www.academicjournals.org

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The African Journal of Food Science (AJFS) (ISSN 1996-0794) is published monthly (one volume per year) by Academic Journals.

African Journal of Food Science (AJFS) provides rapid publication of articles in all areas of Food Science such as Sensory analysis, Molecular gastronomy, Food safety, Food technology etc. The Journal welcomes the submission of manuscripts that meet the general criteria of significance and scientific excellence. Papers will be published shortly after acceptance. All articles published in AJFS are peer-reviewed.

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Dr. Thaddeus Chukwuemeka Ezeji Dr. Hu Xiao-Qing

Ohio State University and State Key Lab of Food Science and Technology,

Ohio State Agricultural and Development Jiangnan University,

Center (OARDC) China.

Department of Animal Sciences USA. Dr. Dominic Agyei Department of Food Science/Te Tari Pütaiao Kai Prof. Kofi E. Aidoo University of Otago, Department of Biological and Biomedical Dunedin, Sciences New Zealand. Glasgow Caledonian University Glasgow Scotland. Dr. Fook Yee Chye Faculty of Food Science and Nutrition, Universiti Malaysia Sabah, Dr. Barakat S.M. Mahmoud Malaysia. Food Safety/Microbiology Experimental Seafood Processing Laboratory Costal Research and Extension Centre Dr. Adel Shatta Mississippi State University Department of Food Technology, USA. Faculty of Agriculture, Egypt.

Dr. Neela Badrie Department of Food Production, Dr. Tendekayi Henry Gadaga Faculty of Science and Agriculture, Departmentof Environmental Health Science University of the West Indies, University of Swaziland Trinidad and Tobago. Swaziland.

Editorial Board Members

Dr. K. Pandima Devi

Department of Biotechnology

Alagappa University

Tamil Nadu India.

Dr. Ashish Kumar Singh Dairy Technology Division National Dairy Research Institute, Haryana, India.

Prof. Rui Cruz Department of Food Engineering Institute of Engineering University of Algarve, Faro Portugal.

African Journal of Food Science

Table of Content: Volume 12 Number 6 June 2018 Issues

ARTICLES

Chemical composition, functional and organoleptic properties of complementary foods formulated from millet, soybean and African locust bean fruit pulp flour blends 126 Zakari U. M., Hassan A. and Fatima Kida

Meat quality analysis of Djallonke lambs co-infected with Teladorsagia circumcincta and Trichostrongylus colubrifromis (Trichostrongyloidae) and treated with two medicinal plants 131 Ndah Germaine, Fonteh A. Florence, Tedonkeng Fernand, Mpoame Mbida, Lumofoet Jules, Temgoua I. F. Kengne , Kenmogne A.Tayou, Nzonang N. Douglas, Vemo B. Narcisse and Wabo J. Poné

Comparison of traditional butter preservation techniques using microbial and organoleptic properties, West Shewa, Ethiopia 140 Alganesh Tola, Debela Bayu, Lemma Fita, Bilatu Agza and Sara Birkie

Vol. 12(6) pp. 126-130, June 2018 DOI: 10.5897/AJFS2018.1692 Article Number: 754612957065 ISSN 1996-0794 Copyright © 2018 Author(s) retain the copyright of this article African Journal of Food Science http://www.academicjournals.org/AJFS

Full Length Research Paper

Chemical composition, functional and organoleptic properties of complementary foods formulated from millet, soybean and African locust bean fruit pulp flour blends

Zakari U. M.1, Hassan A.2* and Fatima Kida 1

1Department of Food Technology, Kaduna Polytechnic, Kaduna, Nigeria. 2Department of Nutrition and Dietetics, Kaduna Polytechnic, Kaduna, Nigeria.

Received 30 January, 2018; Accepted 28 March, 2018

This study was aimed at producing and evaluating the quality of complementary foods from millet, soybean and African locust bean fruit pulp. The millet, soybean and African locust bean fruits were obtained locally in Kaduna, Nigeria. These materials were separately cleaned, dried, milled and sieved into flours. The flours were mixed as follows: Sample A (60% millet, 10% soybean, 30% African locust bean fruit pulp), sample B (60% millet, 20% soybean, 20% African locust bean fruit pulp), Sample C (50% millet, 40% soybean, 10% African locust bean fruit pulp), sample D (50% millet, 30% soybean, 20% African locust bean fruit pulp), sample E (50% millet, 20% soybean, 30% African locust bean fruit pulp). The chemical composition, functional properties and sensory attributes were determined using standard methods. The moisture content of the food samples varied from 7.5 to 11%, protein ranged from 28.9 to 37.0%, fat from 3.1 to 5.1%, Ash ranged from 1.4 to 2.51%, crude fibre ranged from 2.0 to 3.2%, carbohydrate varied ranged from 47.0 to 52.1%. The energy values ranged from 351 to 381.9 kcal/g and appreciable amount of minerals were recorded. Calcium ranged from 128.0 to 165.0 mg/100 g, iron ranged from 3.2 to 5.8 mg/100 g and zinc ranged from 2.6 to 3.2 mg/100 g. The results of functional properties showed that bulk density ranged from 0.61 to 0.66 g/ml, water absorption capacity ranged from 1.89 to 2.31 g/g, swelling capacity ranged from 0.67 to 2.45% and least gelation ranged from 9.00 to 11.62%. Sensory evaluation indicated that the samples were highly rated (P<0.05) for all the parameters investigated. Sample E (50% millet, 20% soybean and 30% African locust bean pulp flour) was most acceptable to the panelists and should be encouraged in weaning food preparation.

Key words: African locust bean fruit pulp, complementary foods, functional oroperties, chemical composition.

INTRODUCTION

Protein-energy malnutrition among children is the major also be irreversible sequence from micronutrient health challenges in developing countries, particularly deficiencies that affect brain development and other Nigeria (FAO, 2001; Ijarotimi and Keshinro 2012). functional outcomes (Martorell et al., 1995). This nutrition Malnutrition during early life leads to permanent stunting problem is associated with inappropriate complementary in growth (Nzeagwu and Nwaejike, 2008) and there may feeding practices, low nutritional quality of traditional

Zakari et al. 127

complementary foods and high cost of quality protein Government of Kaduna State. All the equipment used for this study based commercial complementary foods (Nemer et al., were from the Department of Food Technology Kaduna 2001; Muller et al., 2003; Black et al., 2003; FAO, 2004, Polytechnic, Kaduna. The chemicals used for analyses were of analytical grade and obtained from the laboratory of the Department Alozie et al., 2009; Eka et al., 2010). The tragic of Food Technology, Kaduna Polytechnic, Kaduna. The chemicals consequences of malnutrition include death, disability, were products of synth, Sao Paulo, Brazil. stunting, mental and physical growth and as a result, retarded national socio-economic development (Ijarotimi and Keshinro, 2012). It is evidence that high prevalence Preparation of materials of deaths each year among children aged under five Soybean seeds were cleaned, sorted and roasted at 100°C for 2 h. years in the developing world are associated with The roasted seeds were cooled and milled into flour using a malnutrition (WHO, 2002). The interaction of poverty, laboratory hammer mill (Christy Hunt, UK). The flour was sieved poor health and poor complementary feeding practices using a 60 mm mesh sieve (British Standard). The flour was packed has a multiplier effect on the general welfare of the in a plastic container, sealed and stored at room temperature children population and contributes significantly towards (25°C). The millet grains were cleaned and sorted. The sorted grains were then milled into powder using the laboratory hammer growth retardation, poor cognitive development, illness mill and sieved to fine powder using a 60 mm mesh sieve. and death among children in developing countries, The locust bean fruit pulp flour was prepared using the method particularly Nigeria (Pollit, 1994; Duncan et al., 1994; described by Zakari et al. (2013) and Zakari et al. (2015). The outer Kretchmer et al., 1996; Bhattacharya et al., 2004; Anigo brown cover of the pods was manually striped open and the yellow et al., 2007). pulp was separated from the seeds embedded within the pulp. The The complementary feeding which usually begins at 4 yellow pulp was dried in a hot air oven (model T121, Gen lab Widnes, UK) at 60°C for 9 h to moisture content of 10.5%. The to 6 months continues up to the age of 24 months when dried powder was milled with a laboratory hammer mill and sieved the transition from exclusive breastfeeding to semi solid using a 60 mm mesh sieve. The flour was also packed in a plastic food begins. It is at this stage that the nutritional container and stored at room temperature (25°C) prior to analyses. requirements of many infants are not met, thus leading to onset of malnutrition that is prevalent in children under 5 years of age (Anigo et al., 2009). The traditional Food formulation complementary foods in Nigeria are cereal based (e.g. Composites were formulated as indicated on table 1, from the ogi) and other family diets (tuwo, cassava, yam, etc.) and processed flours in ratio of 60:10:30, 60:20:20, 60:30:10, 50:30:20, these plant based complementary foods are not 50:20:30 of millet, soybean and locust bean in pulp, to obtain beneficial to the growth and development of children products A, B, C, D and E, respectively. (Ijarotimi and Keshinro, 2012). Investigations have shown that ogi (corn gruel, a traditional complementary food) and other family diets often fail to meet the nutritional Analytical determinations needs of the infants due to poor nutritive values The nutrient composition of the flour samples was determined (Fernandex et al., 2002; Solomon, 2005,), hence they according to the standard assay methods of AOAC (2005). have been implicated in the etiology of protein energy Moisture content was determined by oven method, crude protein by malnutrition in the community where they are solely used microkjedahl method using 6.25 as a conversion factor. Fat was as complementary foods (Okoye, 1992, Devlin, 1997). determined by ether extraction using soxhlet extractor and ash was In view of the nutritional problem associated with the determined by drying ashing method. The carbohydrate content was determined by difference. Addition of all the percentages of traditional complementary foods, coupled with high cost moisture, fat, crude protein ash and crude fibre was subtracted from of commercial baby foods, this study is therefore aimed 100%. This gives the amount of nitrogen free extract otherwise at formulating complementary foods from millet, soybean known as carbohydrate. All the analyses were determined following and African locust bean fruit pulp (Parkia biglobosa) and the method of AOAC (2005): to evaluate the quality of the formulated foods. % Carbohydrate = 100 – (% moisture + % fat + % Ash + % Crude fibre + % crude protein).

MATERIALS AND METHODS The sample calorific value was estimated (in kcal/g) by multiplying the percentages of crude protein, fat and carbohydrate using Soybeans and millet grains were purchased from Monday Market in Atwater conversion factor (4, 9, and 4, respectively) as proposed by Kaduna South Local Government, Kaduna. Locust bean fruits were Mahan and Escott-Stump (2008). Calcium, Iron and Zinc were bought from Kasuwan Magani, a village in Kajuru Local determined by atomic absorption spectrophotometer.

*Corresponding author. E-mail: [email protected]

Author(s) agree that this article remain permanently open access under the terms of the Creative Commons Attribution License 4.0 International License

128 Afr. J. Food Sci.

Table 1. Recipe formulation.

Samples Component A B C D E Millet flour 60 60 50 50 50 Soybean flour 10 20 40 30 20 African locust bean fruit pulp 30 20 10 20 30

Functional properties of the flour samples were also determined. water for microbial activity is low. This report agreed with Bulk density, water absorption capacity were determined by Okaka similar work of Ijarotimi and Keshinro (2012). and Porter (1979). The gelation properties were determined by the The protein content ranged from 28.9 to 37%. The high method of Coffman and Garcia (1977). The swelling capacity was determined by Coffman and Garcia, (1977) with slight modifications. levels of protein could be a result of supplementation with One gram of the flour sample was mixed with 10ml distilled water in soybean. The protein levels increased with increasing a centrifuge tube and heated at 80°C for 30 min. This was addition of soybean flour. Nnam (2001) made a similar continually shaken during the heating period. After heating, the observation when the sorghum traditional complementary suspension was centrifuged at 1000 × g for 15 min. The food was supplemented with bambara groundnut and supernatant was decanted and the weight of the paste taken. Swelling capacity was calculated as weight of the paste/weight of sweet potato. Similarly, Nzeagwu and Nwaejike (2008) dry flour. reported increases in protein when sorghum traditional complementary food was supplemented with groundnut and crayfish. The content also follows the same trend of Sensory evaluation increases with increasing addition of soybean. The fat varied from 3.00 to 5.1%. The high fat content of soybean Porridges were prepared from each of the composite flours. Briefly, 20 g of each of the samples were homogenized in 60 ml of water resulted in the high fat level of sample C (50:40:10). The and the slurry was heated slowly with constant stirring for 15 min, ash content varied from 1.4 to 2.51%. The carbohydrate one teaspoon of sugar was added to each sample. The porridges content ranged between 47.0 to 52.1%. The energy value were kept separately in thermos flask for sensory evaluation. ranged from 351 to 381.9 kcal with sample C having the Sensory evaluation was conducted on the reconstituted samples highest energy value. The energy values of the formulated which were coded and presented to 15 untrained panelists. The samples met the FAO/WHO (1991) specification sensory evaluation was conducted in a well standard sensory evaluation room, where each of the panelists was positioned in a guidelines for young children complementary food separate cubicle to avoid interference. The samples were rated on formulations. The mineral composition of the formulated the following sensory characteristics, that is, colour, taste, food samples shown on Table 2, indicated that calcium mouthfeel and overall acceptability using 9-point hedonic scale, ranged from 128 to 165 mg/100g, with sample A having with 9 as like extremely and 1 as dislike extremely (Ihenkoronye the highest calcium level. The high calcium levels in and Ngodi, 1985). sample A may be due to high proportion of millet and African locust bean fruit pulp flour. The iron and zinc Statistical analysis content were also higher in sample A (60:10:30), with the lowest percentage of soybean. The data obtained from the study were analyzed using means and The functional properties of the formulated food standard deviation. Analysis of Variance (ANOVA) and Duncan’s materials are shown in Table 3. The results showed that New Multiple Range Test (DNMT) (Ihekoronye, 1985) were used to test the significance between the means (p<0.05) of sensory bulk density ranged from 0.61 to 0.66 g/ml, water scores. absorption capacity ranged from 1.89 to 2.31% swelling capacity ranged from 0.67 to 2.45%, least gellation ranged from 9.0 to 11.62%. The result showed that the RESULTS AND DISCUSSION samples possess low bulk density, which is an advantage in the preparation of complementary food, because high Table 2 presents the nutrient composition of millet, bulk limits the caloric and nutrient intake per feed of a soybean and African locust bean fruit pulp flour blends. child (Onimawo and Egbekun, 1998; Omueti et al., 2009). The moisture content of blends ranged from 7.5 to The low bulk density is economical in food packaging. 11.0%. The moisture content of the blends (7.5 to 11.0%) The water absorption capacity is also at lower level for all was within the range for flour (11 to 12%) as reported by the samples. The water absorption capacity is an index of Ihekoronye and Ngoddy, 1985). This moisture level is the maximum amount of water that a food product would also in agreement with the work of Nzeagwu and absorb and retain (Ijarotimi and Keshinro, 2012). With Nwaejike (2008). The low moisture content is desirable respect to water absorption capacity, Giami and as it enhances the keeping quality of the samples since Bekehand (1992) reported that microbial activities of food

Zakari et al. 129

Table 2. Proximate and mineral composition of millet, soybean and locust bean fruit pulp blends.

Sample A Sample B Sample C Sample D Sample E Parameter (60:10:30) (60:20:20) (50:40:10) (50:30:20) (50:20:30) Moisture (%) 10.5 ± 0.14a 10.5 ± 0.14a 7.5 ± 0.21c 8.0 ± 0.14b 11.0 ± 0.49a Protein (%) 28.9 ± 0.14d 32.7 ± 0.14c 37 ± 0.20a 35 ± 0.02b 31.6 ± 0.21c Fat (%) 3.00 ± 0.20d 4.5 ± 0.02b 5.1 ± 0.02a 4,8 ± 0.20c 4.46 ± 0.20c Ash % 2.5 ± 0.02a 2.0 ± 0.02b 1.4 ± 0.14c 2.1 ± 0.14b 2.51 ± 0.02a Crude Fibre (%) 3.00 ± 0.02a 2.5 ± 0.02b 2.0 ± 0.02c 2.51 ± 0.02b 3.2 ± 0.02a Carbohydrate (%) 52.10 ± 0.22a 47.80 ± 2.00b 47.0 ± 0.02e 47.59 ± 0.02c 47.2 ± 0.02e Energy (Kcal/g) 351c 362.5c 381.9a 373.56b 355.61d Calcium (mg/100g) 165 ± 0.2a 160 ± 0.2c 128 ± 0.4e 159 ± 0.1 163 ± 0.2b Iron (mg/100g) 5.8 ± 0.2a 4.8 ± 0.1b 3.2 ± 0.2d 4.7 ± 0.2b 4.7 ± 0.2c Zinc (mg/100g) 3.2 ± 0.01a 2.6 ± 0.02d 2.7 ± 0.01c 2.9 ± 0.2b 2.93 ± 0.2b

Means with different superscripts on horizontal line are significantly (P<0.05) different.

Table 3. Functional properties of millet, soybean and locust bean fruit pulp blends.

Sample A Sample B Sample C Sample D Sample E Parameter (60:10:30) (60:20:20) (50:40:10) (50:30:20) (50:20:30) Bulk Density (g/ml) 0.65 ± 0.05a 0.60 ± 0.05c 0.63 ± 0.02b 0.6 ± 0.02c 0.65 ± 0.05a Water absorption capacity (g/g) 2.19 ± 0.2c 1.96 ± 0.02d 2.04 ± 0.02b 1.89 ± 0.2e 2.31 ± 0.02a Swelling capacity (%) 0.67 ± 0.04c 2.22 ± 0.03d 2.45 ± 0.02a 2.42 ± 0.03b 2.41 ± 0.02c Least Gelation (%) 9.00 ± 0.02d 11.50 ± 0.04c 11.62 ± 0.01b 11.61 ± 0.03b 11.51 ± 0.02c

Means with different superscripts on horizontal line are significantly (P<0.05) different.

Table 4. Sensory attributes of complementary foods from millet, soybean and African locust bean fruit pulp (P=0.05).

Sample A Sample B Sample C Sample D Sample E Parameter (60:10:30) (60:20:20) (50:40:10) (50:30:20) (50:20:30) Taste 6.87c 7.23c 6.73c 7.03c 7.93a Mouthfeel 7.00c 7.46b 6.86c 7.46b 7.93a Colour 6.40a 6.0a 7.00C 7.60a 7.80a Overall acc 6.93a 7.33c 6.80d 7.60b 7.93a

Means with different superscripts on horizontal line are significantly (P<0.05) different.

products with low water absorption capacity would be soybean and 30% African locust bean fruit pulp was more reduced. Hence the shelf life of such product would be acceptable to the panelists on all the attributes tested. extended. The swelling capacity is used in the determination of the amount of water that food samples can absorb and the degree of swelling within a given Conclusions time. The gelling properties indicated that the products can be used as complementary food formulation. The study has shown that acceptable complementary The results of sensory evaluation are shown in table 4. food can be produced from composites of millet, soybean The results showed that there are significant (<0.05) and African locust bean fruit pulp. This study has also differences in all the attributes tested for the samples. shown that composites of millet, soybean and African However sample E (50.20:30) with 50% millet, 20% locust bean fruit pulp are nutritionally adequate and

130 Afr. J. Food Sci.

Mahan LK, Escott-Stump S (2008). Krause’s Food & Nutrition Therapy, possess good functional and sensory properties, which th are required for the preparation of complementary foods 12 ed. Sauders Elsevier, Canada, pp. 46-57 Martorell R, Schroeder DG, Rivera JA, Kaplowitz HJ (1995). Patterns of for infants and children. linear Italics growth in rural Guatemalar adolescent and children. J. Nutr. 125(supple):10605-10675. Muller O, Garenne M, Kouyate B , Becher H (2003). The association CONFLICT OF INTERESTS between protein-energy malnutrition, malaria morbidity and all-cause mortality in West African children. Trop. med Int. Health 8:507-511. Nemer L, Gelband H, Jha P (2001). Commission on Macroeconomics The authors have not declared any conflict of interests. and Health. The evidence based for interventions to reduce malnutrition in children under five and school-age children in low and middle-income countries. CMH working paper No. WG5:11. REFERENCES Geneva: World Health Organisation, 2001. Nnam NM (2001). Chemical, Sensory and rheological properties of Alozie YE, Iyam MA, Lawal O, Udofia U, Ani IF (2009). Utilization of porridges from processed sorghum, bambara groundnut and sweet Bambara groundnut flour blends in bread production. J. Food potatoes flours. Plant Food, Hum. Nutr. 56:251-264. Technol. 7(4):111-114. Nzeagwu OC, Nwaejike NJ (2008). Nutrient Composition, functional and Anigo KM, Ameh DA, Ibrahim S, Danbauchi SS (2007). Microbiological organoleptic properties of complementary foods formulated from analyses of Commonly used local complementary foods in North sorghum, groundnut and crayfish. Nigeria Food J. 26:13-20. Western Nigeria. J. Med. Sci. (5):875-879. Okaka JC, Potter NN (1979). Physicochemical and functional properties Anigo KM, Ameh DA, Ibrahim S, Danbauchi SS (2009). Infant feeding of cowpea podres processed to reduce beany flavor. J. Food Sci. practices and nutritional status of children in North Western Nigeria. 44:1235-6240. Asian J. Clin. Nutr. (1):12-22. Okoye ZSC (1992). Biochemical Aspects of Nutrition Prentice-Hall of Association of Official Analytical Chemists (AOAC) (2005). Official India, New Delhi, pp. 147-150. methods of Analysis of the Association of Analytical Chemists, Ijarotimi OS, Keshinro OO (2012). Formulation and Nutritional Quality International, 18th ed. Gathersburg, M.D. U.S.A. Official methods. of Infant Formula produced from germinated popcorn, Bambara Black RE, Morris SS, Bryce J (2003). Where and why are 10 million groundnut and African locust bean flour. J. Microbiol. Biotechnol. Children dying every year? In Lancet (361):226-234. Food Sci. 1(6):1358-1388. Bhattacharya J, Currie J, Haider S (2004). Poverty, food insecurity and Onimawo AI, Egbekun KM (1998). Comprehensive Science and nutritional outcomes in children and adults. J. Health Econ. (28):839- nutrition, Ambik Press Ltd. Benin City pp. 103-208. 862. Omueti O, Otegbayo B, Jaiyeola O, Afolabi O (2009). Functional Coffman CW, Garcia VV (1977). Functional Properties and Amino Properties of Complementary diets developed from soybean, acid content of a protein isolate from mug bean flour. J. Food groundnut and crayfish. In EJEAFChe, 8(8):563-573. Technol. (12):473-484. Pollit E (1994). Poverty and Child Development: Relevance of Research Devlin MT (1997). Textbook of biochemistry with Clinical Correlations. in Developing Countries to the United States. In Child Development 4th ed. Wiley-Liss In. New York/Chichester/Toronto. pp. 540-548. Countries to the United States. In Child Development. 64(2):283-295. Duncan JG, Brooks-Ciunn J, Klebanov PK (1994). “Economic Solomon M (2005). Nutritive Value of three Potential Complementary Deprivation and Early Childhood Development”. Child Dev. Foods based on Cereals and legumes. Afr. J. Food Nutr. Sci. 5(2):1- 65(2):296-318. 14. Eka BE, Abbey BW, Akaninwo JO (2010). Nutritional Evaluation of World Health Organization (WHO) (2002). Childhood Nutrition and Some Traditional Weaning Foods from Akwa Ibom State, Nigeria. In Progress in Implementing the International Code of Marketing of Nigerian. J. Biochem. Mol. Biol. 25(1):65-72. Breast-milk substitutes. Geneva, World Health Organization, 2002 Food and Agricultural Organization (FAO) (2001). Targeting for nutrition (document A55/14). resource for advancing wellbeing. Rome: Food and Agricultural Zakari UM, Ajayi SA, Hassan A (2013). Effect of locust bean fruit pulp Organization, U.N. Geneva. (Parkia biglobosa) flour on the quality of wheat biscuits. J. Agri. Food and Agricultural Organization (FAO) (2004). Undernourishment Biotechnol. Sustain. Dev. 5(6):99-102. around the world. In: The taste of food insecurity in the world. Zakari UM, Hassan A, Ndife J (2015). Quality characteristics of African Food and Agricultural Organization/ World Health Organization locust bean fruit pulp cakes. Afr. J. Food Sci. 9(1):17-22. FAO/WHO (1991). Codex, CAC/GL. Codex Alimentarius. Guidelines on Formulated Supplementary Foods for Older Infants and Young Children.FAO/WHO joint Publications. P 144. Fernandex DE, Vanderjagt DJ, Williams M, Huarg YS, Lut-Te C, Millson M, Adnrew R, Pastuszyn A, Glew RH (2002). Fatty acids, amino acids, and trace mineral analyses of five weaning foods from Jos, Nigeria. Plant Foods Human Nutr. (57):257-274. Giami SY, Bexham DA (1992). Proximate Comositionand functional properties of raw and processed full fat fluted pumpkin (Telferia occidentalis) seed flour. J. Food Sci. Agric. (59):32. Ihekoronye AI, Ngoddy PO (1985). Integrated Food Science and Technology for the Tropics, Macmuillan Publishers London. Kretchmer N, Beard JL, Carlson S (1996). “The role of Nutrition in the Development of Normal Cognition”. Am. J. Clin. Nutr. (63):997-1001.

Vol. 12(6) pp. 131-139, June 2018 DOI: 10.5897/AJFS2018.1690 Article Number: 42B814357073 ISSN 1996-0794 Copyright © 2018 Author(s) retain the copyright of this article African Journal of Food Science http://www.academicjournals.org/AJFS

Full Length Research Paper

Meat quality analysis of Djallonke lambs co-infected with Teladorsagia circumcincta and Trichostrongylus colubrifromis (Trichostrongyloidae) and treated with two medicinal plants

Ndah Germaine1, Fonteh A. Florence2, Tedonkeng Fernand2, Mpoame Mbida1, Lumofoet Jules2, Temgoua I. F. Kengne 2, Kenmogne A.Tayou 2, Nzonang N. Douglas2, Vemo B. Narcisse2 and Wabo J. Poné 1

1Research Unit of Biology and Applied Ecology, Faculty of Science, University of Dschang, Cameroon. 2Department of Animal Production, Faculty of Agronomy and Agricultural Sciences, University of Dschang, Cameroon.

Received 19 January, 2018; Accepted 20 March, 2018

Gastrointestinal nematode infection adversely affects small ruminant productivity all over the world, especially in tropical countries. A study was carried out to evaluate the extent to which concurrent infection with these nematodes may influence lamb meat quality, and how phytotherapy might improve these parameters. During the trial, 24 male Djallonke lambs (Age: 3 to 5 months) were experimentally infected with Teladorsagia circumcincta (4000 infective larvae) and Trichostromgylus colubriformis (10000 infective larvae) and treated from day 26 post infection with methanol extracts of Harungana madagascariensis and Momordica foetida twice daily. Lambs were divided into 8 groups: An untreated (Group 1), Albendazole 7.5 mg/kg (Group 2), H. madagascariensis extract at 125 mg/kg (Group 3), H. madagascariensis extract at 250 mg/kg (Group 4), H. madagascariensis at 500 mg/kg (Group 5), M. foetida extract at 125 mg/kg (Group 6), M. foetida 250 mg/kg (Group 7), and M. foetida at 500 mg/kg (Group 8). Meat quality attributes of lambs from different treatment groups were evaluated. Treatment with different doses of plant extracts led to differences in the level of parasitic load at necropsy. This effect was accompanied by an elevated pH24 in meat from the most severely infected lambs and a concomitant decrease in their chemical composition. This high pH24 equally had significant influence on meat flavour, overall acceptability and water retention of lamb meat. The present study suggested that nematode infection may influence pH of resulting carcass, nutritional composition and some sensory parameters of meat while treatment using M. foetida at 500 mg/kg maybe important in reducing parasite burden while enhancing quality of subsequent lamb carcass.

Key words: Phytotherapy, meat quality, Teladorsagia circumcincta, Trichostrongylus colubriformis, Harungana madagascareinsis, Momordica foetida, Djallonke lambs.

132 Afr. J. Food Sci.

INTRODUCTION

Infection with gastrointestinal nematodes has a worldwide compounds, especially tannins, which show direct and distribution, occurring in both tropical and subtropical indirect anthelmintic activity for gastrointestinal nematode areas of the world (Biu et al., 2012b; Mohamed et al., control in ruminants (Vargas-Magaña et al., 2014; Zhong 2016). In Cameroon, gastrointestinal helminth infections et al., 2015). are very prevalent, especially in the Western High Lands Momordica foetida (Cucurbitaceae) and Harungana of the country where the rainfall pattern and ambient madagascariensis (Hypericaceae) are two medicinal temperature favour the growth and multiplication of plants widely known for their anthelmintic and appetite parasite (Ndamukong et al., 1989). Field trials carried out stimulating potentials (Pavan et al., 2013; Olukayode et by Mbafor et al. (2014) in West Region of Cameroon al., 2008). Boronkini et al. (2012) carried out a indicated that Trichostrongylus and Teladorsagia species phytochemical analysis of both plants and reported that are among the most common nematodes of sheep in the they are rich in secondary metabolites such as tannins, Western Highlands, with a high prevalence of 93.3 and flavonoids and alkaloids. In addition, M. foetida contains 53.3%, respectively. During primary infection with lipids and lipophilic compounds which have been used in Teladorsagia spp., dysplasia of the abomasal glands and fattening domestic herbivores (Oloyede and Aluko, 2012). a reduction in acid-secreting cells occur as the Furthermore, giving ruminants polyphenolic plants could nematodes colonise the stomach mucosa (Fox, 1997; improve meat quality by increasing antioxidants into the Rinaldi et al., 2011). Severe villus atrophy (decreased circulatory system, which when assimilated and retained villous: crypt ratios), goblet cell hyperplasia as well as in the meat, they will improve sensory characteristics sloughing of enterocytes into the intestinal lumen occur such as flavor and juiciness scores of meat (Qwele et al., with establishment of Trichostromgylus colubriformis 2013; Moyo, et al., 2014; Zhong et al., 2015). larvae (Roy et al., 2004). Characteristic dark foul smelling It is clear that infection with T. circumcincta and T. diarrhoea, anorexia, diminished absorptive capacity or colubriformis may depress appetite, reduce nutrient poor food conversion efficiency are the consequences of utilization and deplete glycogen reserves, thus the destruction of the mucosal architecture (Williams and influencing meat quality. However, the extent to which Palme, 2012; Rajib et al., 2014). It has been reported that these nematodes can influence meat quality remains co-infections with Teladorsagia and Trichostrongylus spp. doubtful (Zhong et al., 2015). Moreover, if this is true, can has a synergistic effect (Steel et al., 1982; Sykes et al., administration of plant extracts improve nutrient utilization 1988). The combined effect of anorexia, malabsorption and the resultant meat quality after slaughter? With these and diarrhoea may result in loss of quantity and quality of questions in mind, the present study was designed to resulting carcass (Mushi et al., 2007). evaluate changes in the nutritional value of meat from Nowadays, consumers are highly interested in the lambs co-infected with T. circumcincta and T. quality of the products they eat, especially when this colubriformis, and how treatment use with methanol refers to meat (Guerrero et al., 2013). Furthermore, meat extracts of M. foetida and H. madagascariensis may quality is of significant importance for consumers' health improve these parameters. (Ivanov et al., 2017). It has been reported that infections with gastrointestinal nematodes affect some meat characteristics, including the meat color stability, METHODOLOGY composition and conformation of small ruminants (Arsenos et al., 2009; Zhong et al., 2015). Study animals

Although, chemical anthelmintics have long been used A total of 24 male Djallonke sheep were used in this study. Ages of in combating helminthiasis, their use is increasingly the animals ranged between 4 and 5 months. Dentition pattern as discredited due to the increasing resistance of these described by Samad (2008) was used to determine ages of nematode to most classes of anthelmintics available, experimental animals. These animals were acquired from two drug residues in animal products and loss of productivity sheep farmers in Fongo Tongo village, 40 km away from Dschang town. Two weeks before arrival, all the animals in the two farms in host animals (Zhong et al., 2015., Jackson and Coop, were vaccinated against Small Ruminant Pests (SRP) using Capri- 2000; Tsiboukis et al., 2013). The general swing towards pestovax and were also treated with anthelmintics (levamisole the use of natural compounds has stimulated research injection 10 mg/kg and Albendazole tablets 10 mg/kg). Two days into their use as anthelminthic alternatives. Thus, many after arrival of animals in the experimental animal house of the studies have explored the use of plant-derived phenolic Faculty of Agronomy and Agricultural Sciences of the University of

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Germaine et al. 133

Dschang, experimental animals were given antibiotics hind feet (at the tarsal-metatarsal joint). The viscera including the (Oxytetracycline 1 ml/10 kg body weight), anticoccodia (Toltrazuril, gastrointestinal tract were removed. 10 mg/kg), multivitamin and anti-stress (Stress vita 2.5 ml/10 kg body weight). They were allowed to acclimatize for 14 days and their faecal samples examined daily within this period to confirm Meat quality analysis that animals were helminth free by the use of the concentrated salt flotation technique as described by Thienpont et al. (1986) and Meat samples from longissimus dorsi muscle were taken between 6 Baker (2007). After this adaptation period, all animals were and 11th ribs of each animal and used for physical and inoculated with the same dose of 14000 L larvae as follows: day 1: 3 technological evaluation while meat from the left hind leg was used 5.000 L3 of T. colubriformis per animal using a syringe via the oral for chemical and sensory evaluation. route. Day 2: 5.000 L3 of T. colubriformis and day 3: 4.000 L3 of T. circumcincta. Animals were randomly allocated to eight sub groups of 3 individuals on day 26 post inoculation and treated as follows: Group 1, Untreated group (received the vechicle 3.2% dimethyl Physical evaluation sulphoxide); Group 2, Positive control group (Albendazole 7.5 mg/kg); Group 3, H. madagascariensis 125 mg/kg; Group 4, H. Determination of pH24 madagascariensis 250 mg/kg; Group 5, H. madagascariensis 500 mg/kg; Group 6, M. foetida 125 mg/kg; Group 7: M. foetida 250 To determine pH24, 10 g of l. dorsi muscle from each carcass was mg/kg; Group 8, M. foetida 500 mg/kg body weight. Each group weighed using an electronic balance (max 6000 g, d = 0.1 g) and 2 was placed in separate pens of dimension 7.5 m (3 m × 2.5) on refrigerated at 0°C for 24 h. The ultimate pH24 was evaluated after slatted floors. A solid partition separated adjacent pens and care thawing by inserting the sharp tip of a portable pH meter (Testo was taken to avoid contamination of pens with nematode larvae 205, Testo AG, Lenzkirch, Germany) to a depth of about 2 cm into from outside sources. All animal groups were served 6 kg of the each meat sample. The pH value was then read directly on the pH same fresh forage (mainly Tripsacum laxum and Leucaena meter. leucocephala) harvested 24 h before feeding, washed in clean water, dried in the sun for 2 h and chopped before serving to the animals. Water was provided ad libitum. The weight of the animals was taken weekly and weight of food consumed by each group was Evaluation of technological properties of meat recorded daily. Drip loss

Selection of plants The drip loss was evaluated following the procedure described by Honikel (1998). One hour after slaughter, 100 g of muscle was An ethnobotanic study permitted the selection of two medicinal weighed and put in nylon net bags. Each bag was hung over a plants with anthelmintic and appetite stimulating potentials. They beaker at 4°C for 24 h, after which the samples were removed, were harvested in April 2016, identified and authenticated at the dabbed dry on a serviette and reweighed. Percentage drip loss was National Herbarium of Cameroon in Yaounde by voucher specimen calculated using the formula: N° 46105/HNC for H. madagascariensis and N° 24393/HNC for M. foetida. The bark of H. madagascariensis and whole aerial parts of Drip loss (%) = (2) M. foetida were shade dried at room temperature. The dried plant (2) parts were powdered mechanically using a commercial electrical blender and stored in airtight plastic bags at 4°C until preparation of extracts. Cook -out loss

The cook-out-loss was evaluated as described by Paisentier et al. Preparation of methanol extracts (2003). To this effect, 50 g of l. dorsi muscle was weighed and put inside weighed Ziploc bags. These were immersed in a thermostatic Four hundred and fifty grams of each plant powder were macerated water bath at 75°C for 15 min, after which the muscles were in 3 L of methanol. The mixture was stirred for 10 min and stored for removed, allowed to cool to room temperature for about 1 h, 72 h at room temperature. This solution was sieved through dabbed on a paper serviette and re-weighed. The percentage cook- Whatman No. 1 filter paper, divided into portions of 200 ml each out -loss from each sample was calculated as follows: then evaporated in a rotavapor at 50°C. The dry extracts were stored at 4°C until used for in vivo assays. Cook-out-loss (%) = (3) (3) Treatment of experimental lambs

Treatment with plant extracts was administered on day 26 post Freezing loss inoculation and subsequently twice daily for 4 days. Albendazole 7.5 mg/kg was administered on the last day of treatment. On day 56 This was determined based on the procedure described by post inoculation (30 days post treatment), all 24 animals were Paisentier et al. (2003). To this effect, 33 g of l. dorsi muscle from humanely sacrificed, the slaughtering was carried out by severing each lamb carcass was weighed into zip lock plastic bags. These the jugular vein and the carotid arteries to ensure proper bleeding were frozen at -20°C for 14 days. They were then removed and (Kefyalew et al., 2013). The lambs were bled within 3 min after allowed to thaw at room temperature, after which each sample was slaughter and suspended to remove their skin, head (at the dabbed dry and re-weighed. Percentage freezing loss was occipito-atlantal joint), forefeet (at the carpal-metacarpal joint), and calculated as follows:

134 Afr. J. Food Sci.

Figure 1. Influence of treatment using methanol extracts of Harungana madagascariensis and Momordica foetida on pH24 of meat from lambs co-infected with Teladorsagia circumcincta and Trichostrongyglus colubriformis. abBars with different letters are significantly different. DMSO: Dimethyl sulphoxide (3.2%); Alb: Albendazole 7.5 mg/kg; H125: Harungana madagascariensis 125 mg/kg; H250: Harungana madagascareinsis 250 mg/kg; H500: Harungana madagascariensis 500 mg/kg; M125: Momordica foetida 125 mg/kg; M250: Momordica foetida 250 mg/kg; M500: Momordica. foetida 500 mg/kg.

Sensory attributes assessed included tenderness, juiciness, flavour intensity, and overall acceptability as follows: (tenderness: 5 = Freezing loss (%) = tender, 1= very tough; juiciness: 5 = juicy, 1= very dry; aroma: 5 = very good flavour, 1 = off flavour; general acceptability: 5 = highly desirable, 1 = extremely poor). Evaluation of chemical composition of lamb meat

Meat samples were minced and dried in a ventilated oven at 105°C Statistical analysis for 20 to 24 h when a constant weight was attained. Moisture content was calculated as the difference in weight before and after Statistical analysis was performed using SPSS version 20.0. To test drying. Crude protein, crude lipid and ash contents were estimated the effects of treatment on studied parameters, one-way analysis of on dry matter basis as according to the AOAC (2005). The results variance (ANOVA), General Linear Model approach, was used. were expressed as percentages. Pearson correlation coefficient was used to establish the relationship among adult worm burden and pH24 of lamb meat. The post hoc test employed for all analysis was Duncan’s Multiple Minerals analysis Range Test. Results obtained were expressed as mean ± standard deviation. Probability values P<0.05, was considered significant. Calcium (Ca), magnesium (Mg), and iron (Fe), were determined using Atomic Absorption Spectrophotometer (GBC atomic absorption spectrophotometer, Australia) according to Pauwels et al. (1992) and AOAC (2005). The values of Mg, Ca, and P were RESULTS determined as mg/g dry weight, while the value of Fe was measured as mg/kg dry weight. Results showing the variation in adult worm burden recovered at necropsy from the abomasums and intestines of lambs co-infected with T. circumcincta and Evaluation of sensory properties T. colubriformis and treated with albendazole and Sensory characteristics of meat were determined after slaughter of methanol extracts of H. madagascariensis and M. foetida animals using 20 test panellists. Test panellist was selected among in Germaine et al. (2017). students of the Department of Animal Production, Faculty of Figure 1 summarizes the influence of treatment using Agronomy and Agricultural Sciences (FASA), University of methanol extracts of H. madagascariensis and M. foetida Dschang. Screening exercises were administered prior to panel on meat ultimate pH (pH ). selection in order to diagnose the most qualified panel members. 24 These included taste identification and taste intensity exercises, Ultimate pH24 in lamb was observed to be significantly using score charts adapted from Abd El-aal and Suliman (2008) lower (P<0.05) in albendazole treated lambs and those and Dimple and Rohanie (2014). After selection of the most treated with M. foetida at dose rate 500 mg/kg when qualified test panellists, samples of meat were sliced into 2.0 cm compared with untreated group. Treatment with H. steaks, put in zip lock bags based on different treatment groups and madagascariensis extracts at all tested doses and M. boiled for 45 min using an electric cooker. The panellists were foetida served with samples of meat in different trays. To evaluate these at doses less than 500 mg/kg had no significant sensory characteristics, a 5-point scale adapted from Abd El-aal influence (P>0.05) on meat pH24 when compared with and Suliman (2008) and Dimple and Rohanie (2014) was used. untreated lambs. There was a strong positive correlation

Germaine et al. 135

Table 1. Influence of treatment using methanol extracts of Harungana madagascariensis and Momordica foetida on technological properties of lamb meat co-infected with Teladorsagia circumcincta and Trichostrongylus colubriformis.

Treatment Cooking loss (%) Freezing loss (%) Drip loss (%) Vechicle (DMSO 3.2%) 11.14 ± 1.44 9.66 ± 0.04 7.17 ± 0.50b Alb7.5 mg/kg 11.33 ± 2.32 10.31 ± 1.26 9.84 ± 2.82a H125 mg/kg 11.31 ± 1.50 9.31 ± 1.43 7.32 ± 1.71b H250 mg/kg 12.16 ± 2.25 9.16 ± 0.68 7.55 ± 0.39b H500 mg/kg 11.18 ± 1.15 9.46 ± 1.02 7.69 ± 3.39b M125 mg/kg 11.70 ± 1.65 9.85 ± 0.79 7.80 ± 2.21b M250 mg/kg 10.51 ± 1.85 9.44 ± 1.88 7.73 ± 0.75 M500 Mg/kg 11.51 ± 0.43 10.05 ± 1.89 8.70 ± 1.74ab P-value 0.56 0.21 0.01

abcMean values followed by the different latters on the same columns are significantly different (P<0.05). DMSO: Dimethyl sulphoxide (3.2%); Alb: Albendazole 7.5 mg/kg; H125: Harungana madagascariensis 125 mg/kg; H250: Harungana madagascareinsis 250 mg/kg; H500: Harungana madagascariensis 500 mg/kg; M125: Momordica foetida 125 mg/kg; M250: Momordica foetida 250 mg/kg; M500: Momordica foetida 500 mg/kg.

Table 1 shows the effects of treatment on technological DISCUSSION properties of lamb carcasses. There was no significant difference between all groups of animals concerning In this study, methanol was used for extraction because cooking loss and freezing loss. However, significant of its ability to extract almost all the chemical components differences (P ≤ 0.05) in drip loss was found among the of plants due to its high polarity index (Eloff, 1998). Since different animal groups, with those treated with M. foetida almost all identified components like tannin, phenol and 500 mg/kg and albendazole allowing drip significantly alkaloids that are active against helminthes are aromatic more (P<0.05) than all other experimental groups. or saturated organic compounds, they are often obtained Influence of treatment using methanol extracts of H. through initial extraction with methanol or ethanol (Vilegs madagascariensis and M. foetida on chemical properties et al., 1997; Khadijah, 2015). of meat is summarized in Table 2. The lipid content The pH of muscle at post mortem is the main indicator showed no significant variation (P>0.05) between the of meat quality at a commercial level (Cvrtila Fleck et al., different animal groups. Protein and ash contents showed 2015). The present results revealed that pH24 differed a reverse trend, being significantly higher in albendazole significantly between treatment groups. It was observed 7.5 mg/kg, H. madagascariensis 500 mg/kg, M. foetida that apart from lambs that received albendazole and M. 250 mg/kg and M. foetida 500 mg/kg treatment groups. foetida 500 mg/kg, lambs in all other treatment groups This significant difference observed in ash content was (M. foetida 125 mg/kg, M. foetida 250 mg/kg, H. more pronounced in the results of individual mineral madagascariensis 125 mg/kg, H. madagascariensis 250 analysis. Meat from lambs treated with albendazole 7.5 mg/kg, H. madagascariensis 500 mg/kg) and the mg/kg, H. madagascariensis 500 mg/kg, M. foetida 250 untreated group had significantly higher pH24 (pH ≥ 6). mg/kg and M. foetida 500 mg/kg recorded significantly Phillips and Wheeler (2008) reported that ultimate pH24 higher, calcium, magnesium and phosphorus contents (that is after the post mortem biochemical changes when compared with the untreated lambs. However, no involving the depletion of muscle glycogen are significant difference (P>0.05) in iron content across the completed) in lamb tends to be higher in animals different groups was recorded during this study. subjected to chronic stress. Reference has already been Influence of treatment using methanol extracts of H. made to the fact that the pH24 of the best quality lamb madagascariensis and M. foetida on sensory meat tends to fall in the pH range of 5.3 to 5.8 (Beriain et characteristics of lambs co-infected with T. circumcincta al., 2000). The higher pH24 could thus be attributed to and T. colubriformis is summarized in Table 3. There was depleted glycogen reserves, owing to the fact that sheep a significantly higher flavour liking of meat from lambs severely infected with T. circuncuncta and T. treated with albendazole and M. foetida 500 mg/kg than colubriformis have reduced feed intake and significantly all other groups. The overall acceptability result closely lower food utilization ability (Steel et al., 1982; Stear et matched that of flavour liking, with meat from albendazole al., 2003; Rinaldi et al., 2011; Germaine et al., 2017). Abd and M. foetida 500 mg/kg treated lambs being the most El-aal and Suliman (2008) had indicated that ultimate pH acceptable. It was noticed in this experiment that meat is affected by factors such as diet and prolonged hunger from all animal groups was equally tender and juicy. before slaughter. Stress has a marked negative impact

136 Afr. J. Food Sci.

Table 2. Influence of treatment using methanol extracts of H. madagascariensis and Momordica foetida on chemical properties of lamb meat, co-infected with Teladorsagia circumcinta and Trichostrongylus colubriformis.

Chemical Treatment composition DMSO (Vechicle) Albendazole Har 125 Har 250 Har 500 Momo 125 Momo 250 Momo 500 P Proteins (g) 19.70 ± 0.59b 21.76 ± 0.18a 19.30 ± 0.23b 20.38 ± 0.14ab 21.33 ± 0.10a 20.70 ± 0.35ab 21.27 ± 0.49a 21.43 ± 0.65a 0.04 Ash(g) 2.08 ± 0.06b 2.90 ± 0.14a 2.04 ± 0.02b 2.17 ± 0.19b 2.82 ± 0.11a 2.11 ± 0.11b 2.69 ± 0.42a 2.75 ± 0.15a 0.00 Lipids (g) 3.82 ± 0.33 3.94 ± 0.72 3.36 ± 0.75 3.97 ± 1.11 3.32 ± 0.67 3.72 ± 0.34 3.80 ± 0.44 3.84 ± 0.67 0.08 Moisture (g) 75.38 ± 0.65a 73.52 ± 0.92b 75.66 ± 0.33a 75.57 ±0.27a 74.48 ± 0.34ab 76.28 ±0.44a 75.14 ± 0.28a 73.61 ± 0.33b 0.03 Ca (mg) 6.73 ± 0.05b 7.28 ± 0.1a 6.8 ± 0.1b 6.73 ± 0.05b 7.33 ± 0.05a 6.76 ± 0.05b 7.06 ± 0.15a 7.16 ± 0.05a 0.05 Mg (mg) 20.33 ± 3.21b 27.33 ± 0.30a 23.13 ± 1.15b 23.00 ± 2.64b 26.13 ± 0.81a 23.60 ± 2.62b 25.73 ± 0.25a 27.93 ± 0.47a 0.00 Fe (mg) 1.72 ± 0.05 1.93 ± 0.25 1.72 ± 0.06 1.73 ± 0.15 1.83 ± 0.09 1.70 ± 0.10 1.76 ± 0.05 1.88 ± 0.10 0.09 Ph (mg) 175.67 ± 13.05b 197.67 ± 2.51a 183.67 ± 22.54ab 192.33 ±11.06a 200.00 ± 5.29a 171.00 ± 3.00b 200.00 ± 1.00a 204.00 ± 1.00a 0.04

a,bcMeans with the same letters on the same colums are not significantly different (P>0.05). DMSO: Dimethyl sulphoxide (3.2%); Alb: Albendazole 7.5 mg/kg; H125: Harungana madagascariensis 125 mg/kg; H250: Harungana madagascareinsis 250 mg/kg; H500: Harungana madagascariensis 500 mg/kg; M125: Momordica foetida 125 mg/kg; M250: Momordica foetida 250 mg/kg; M500: Momordica foetida 500 mg/kg.

not only on meat pH, but concomitantly on was observed during our investigations that there between 21.5 and 22.5% and moisture content is technological properties of resulting carcass. High was no significant difference between all groups approximately 73.5%. The high moisture content pH24 affects the water binding nature of the of animals concerning cooking loss and freezing could be attributed to increase level of stress due proteins and therefore directly affects the water loss. Our results are in conformity with those to gastrointestinal parasitaemia. Page et al. holding capacity of meat (Huff-Lonergan et al., obtained by Abd El-aal and Suliman (2008) and (2001) reported that stress promotes 2010). During this study, it was observed that Fonteh et al. (2015) who equally recorded no accumulation of intracellular water which reflects meat from lambs in the untreated group had an significant difference in cooking loss and freeing less light and causes the muscle to appear dark. unusually higher water holding capacity, this being loss of meat in groups of animals exposed to On the other hand, the lower protein content could evident by their lower drip loss than albendazole different stress levels. It was observed in this trial be attributed to their significantly lower ability of and M. foetida 500 mg/kg treated lambs. These that carcass from different treatment groups with feed utilization when compared with healthy results are in agreement with the report of Cheng different levels of gastrointestinal adult worm animals. Anorexia and and/or reduction in food et al. (2008) who indicated that high pH meat burden had significantly different levels of conversion efficiency has been observed in sheep does not readily allow drip, thus resulting to dry, moisture and crude protein while the lipid content co-infected with T. colubriformis and T. firm and dark meat. Again, Phillips and Wheeler showed no significant variation. Meat from lambs circumcincta (Steel et al., 1982; Stear et al., 2003; (2008) recorded high water holding capacity treated with plant extract at doses less than 500 Rinaldi et al., 2011; Germaine et al., 2017). corresponding to carcass with the highest pH. mg/kg had a higher water content (>75 g) and Few studies have compared the chemical This high water holding capacity has a major asignificantly lower protein content (<20 g) per composition of meat from lambs with different disadvantage of retaining more water than 100 g of meat. These values deviated from the levels of gastrointestinal nematode parasitism. necessary, leading to more rapid deterioration due standards reported by Sinclair et al. (1990) and However, the present study report agrees with to microbial growth; hence, such meat will have a Williams et al. (2006). These authors indicated Valieva et al. (2014) who observed significantly shorter shelf life (Huff-Lonergan et al., 2010). It that the protein content of 100 g of lamb meat is higher protein and fat content in healthy

Germaine et al. 137

Table 3. Effect of treatment using methanol extracts of Momordica foetida and Harugngana madagascariensis on sensory characteristics of lamb co-infection with Teladorsagia circugmcincta and Harungana madagascariensis.

Treatment Tenderness Juiciness Flavour General acceptability Vechicle (DMSO) 2.67 ± 1.55 3.88 ± 0.90 2.68 ± 0.58b 3.00 ± 0.78c Alb 7.5mg/kg 3.3 ± 1.02 3.67 ± 0.58 4.33 ± 0.00a 4.50 ± 0.00a H125 mg/kg 2.67 ± 0.58 3.0 ± 1.73 2.67 ± 0.58b 3.00 ± 1.00c H250 mg/kg 2.67 ± 0.57 4.00 ± 1.00 2.67 ± 0.58b 3.33 ± 0.58c H500 mg/kg 3.33 ± 1.15 4.33 ± 0.58 3.0 ± 1.0ab 3.67 ± 0.58ab M125 mg/kg 2.68 ± 1.52 3.33 ± 0.57 2.33 ± 0.58b 3.33 ± 0.58c M250 mg/kg 3.0 ± 0.00 4.33 ± 0.57 3.67 ± 0.58a 3.33 ± 1.15c M500 mg/kg 4.33 ± 0.58 4.33 ± 0.58 4.00± 0.00a 4.33 ± 0.58a P-value 0.48 0.53 0.02 0.03

a,bcMeans with the same letters on the same colums are not significantly different (P>0.05). DMSO: Dimethyl sulphoxide (3.2%); Alb: Albendazole 7.5 mg/kg; H125: Harungana madagascariensis 125 mg/kg; H250: Harungana madagascareinsis 250 mg/kg; H500: Harungana madagascariensis 500 mg/kg; M125: Momordica foetida 125 mg/kg; M250: Momordica foetida 250 mg/kg; M500: Momordica foetida 500 mg/kg.

sheep as compared to those suffering from acceptability, with high pH carcass producing undesirable echinococcosis. Again, Sykes et al. (1988) observed that meat with slightly soapy off flavor (Miller, 2007). The most infection with nematode parasites have an adverse effect dominant sulphur (S) compound in cooking meat volatiles upon nitrogen retention, a feature that has been is H2S (Young et al., 1993). These authors further confirmed by the lower protein content of carcass from explained that as meat pH increases, more H2S is sheep co-infected with T. colubriformis or T. circumcincta evolved during cooking. Their data indicate that a pH than in pair-fed controls. This reduced digestibility increase from 5.6 to 6.6, for example, would increase the contributes to the poorer overall economy of such evolution of H2S by about 60%. H2S has its own carcasses with lower protein content. The latter is odourand is also involved in a reaction that generates the primarily a reflection of the lower efficiency with which volatile and very odorous compound thiophenol (benzene they utilize apparently digested nitrogen. Furthermore, thiol). Ha and Lindsay (1991) and Lopez and Lindsay Bermingham et al. (2007) had reported that more skeletal (1993a) showed that thiophenol is the source of muscle protein may need to be degraded and mobilized unpleasant flavour in cooking sheep meat, because to supply increased amino acid demands of the intestine thiophenol's precursor, phenylphosphate, is almost and liver in sheep infected with T. colubriformis. unique to sheep. Thus, either directly or indirectly, high The ash and mineral contents varied between the pH meat might result in enhanced off flavours, hence less different treatment groups. Lambs treated with desirable by test panelists. albendazole, H. madagascariensis 500 mg/kg, M. foetida 500 mg/kg, and M. foetida 250 mg/kg (with lower worm burden) had significantly higher calcium, magnesium and Conclusion phosphorus hence overall ash content. Knox et al. (2006) suggested that the metabolism of mineral ions is modified Parasitism due to co-infection with T. circumcincta and T. by the presence of gastrointestinal nematodes. Sykes colubriformis had a significant influence on meat pH24 and Coop (1976) supported this fact by indicating that with meat from most severely infected lambs (lambs with skeletal calcium and phosphorus in meat from severely the highest worm burden) showing the highest pH24. This infected lambs may be 15 to 45% lower than in high pH24 had significant influence on meat flavour, uninfected animals. overall acceptability and water retention ability of lamb Many consumers judge meat quality from its sensory meat. Furthermore, severely infected lambs were inferior characteristics. In this study, co-infection with T. in chemical composition, having lower protein and circumcincta and T. colubriformis had no significant effect mineral content than healthy lambs, while treatment with on carcass tenderness and juiciness. However, methanol extracts of M. foetida at 250 and 500 mg/kg significant differences were recorded for flavor andoverall and H. madagascariensis at 500 mg/kg led to an acceptability. The most preferred carcasses were from improvement in meat chemical composition. However, albendazole and M. foetida 500 mg/kg treated lambs. further studies are needed to evaluate the muscle energy This could be attributed to pH differences after slaughter. status or lactic acid concentration in the muscles of lambs Carcass pH has a significant effect on flavor and overall with different levels of gastrointestinal parasitism in order

138 Afr. J. Food Sci.

to confirm these results. meat of sheep fed with fed with Agaricus bisporus supplement supplement. Vet. Arhives 85:591-600. Dimple Singh-Ackbarali., Rohanie Maharaj. (2014). Sensory Evaluation as a Tool in Determining Acceptability of Innovative Products CONFLICT OF INTERESTS Developed by Undergraduate Students in Food Science and Technology at The University of Trinidad and Tobago. J. Curriculum The authors have not declared any conflict of interests. Teaching 3(1). Eloff JN (1998). Which extractant should be used for the screening and isolation of antimicrobial components from plants? J. Ethnopharmacol. 60:1-8. ACKNOWLEDGEMENTS Fonteh FA, Bawe NM, Tuncha NP, Bessong OW, Muluh ME, Piasentier E, (2015). Influence of production system on the quality of Gudali beef in Cameroon. Livest. Res. Rural Dev. 27: 29. The authors are grateful to all the sheep farmers and Fox MT (1997). Pathophysiology of infection with gastrointestinal traditional healers in Menoua Division who gave them nematodes in domestic ruminants: Recent developments. Vet. valuable information concerning the plant species they Parasitol. 72:285-308. use as traditional anthelmintic and the methods of Germaine Ndah, Fonteh A, Florence 2, Cedric Yamssi 1, Wabo Poné J (2017). Phytotherapy of Djallonke Lambs Co-infected with preparing and administration of each plant species. Teladorsagia circumcincta and Trichostrongylus colubriformis Further appreciation goes to the Laboratory of Animal (Trichostrongylidae) Using Methanol Extracts of Two Medicinal Plants Biology and Applied Ecology, Laboratory of Animal in Menoua Division, West Region of Cameroon. Euro. J. Med. Plants. Production and Animal Health, Laboratory of Soil 21(3). Guerrero A, VVM, Campo MM, Sañudo C (2013). Some factors that Sciences, all second and third year students of the affect ruminant meat quality: from the farm to the fork. Review. Acta Department of Animal Production in the University of Scientiarum. Anim. Sci. 35(4):335-347. Dschang for providing the environment, resources and Ha JK, Lindsay RC (1991). Volatile alkylphenols and thiophenol in expertise that made this work successful. species-related characterizing flavors of red meats. J. Food Sci. 56:1197-1202. Honikel KO (1998). Reference Methods for the Assessment of Physical Characteristics in Meat. Meat Sci. 49:447-457. REFERENCES Huff-Lonergan E, Zhang W, Lonergan SM (2010). Biochemistry of postmortem muscle-Lessons on mechanisms of meat tenderization. Abd El-aal HA, Suliman AIA (2008). Carcass traits and meat quality of Meat science. 86(1):184-195. lamb fed on ration containing different levels of leucaena hay Ivanov N, Angelova T, Laleva S, Ribarski S, Miteva D, Yordanova D, (Leucaena leucocephala L.). Biotechnol. Anim. Husb. 24(3-4):77-92. Karabashev V, Penchev I (2017). Carcass characteristics and Analysis of the Association of Official Analytical Chemists (AOAC) technological properties of Musculus Longissimus Lumborum at (2005). Official Methods of Analysis of the Association of Official lambs from the Bulgarian dairy synthetic population and its F1 Analytical Chemists 18thEd., AOAC INTERNATIONAL, Gaithersburg, crosses with meat breeds. Agric. Sci. Technol. P 9. MD, USA, Official Method 2005.08 Jackson F, Coop RL (2000). The development of anthelmintic Arsenos G, Fortomaris P, Papadopoulos E, Kufidis D, Stamataris C, resistance in sheep Zygoyiannis D (2009). Meat quality of lambs of indigenous dairy nematodes. Parasitol. 120:95-107. Greek breeds as influenced by dietary protein and gastrointestinal Kefyalew B, Sandip B, Yigrem S (2013). Carcass Traits of Arsi-bale nematode challenge. Meat Sci. 76:779-786. Sheep and Goat Reared under Farmers Management System in Baker DG (2007). Flynn’s parasites of laboratory animals.2nded. Sidama Region of Southern Ethiopia. Middle-East J. Sci. Res. American College of Laboratory Animal Medicine.Blackwel 13(11):1465-1470. publishing. UK. 789p. Khadijah S (2015). Anthelmintic Efficacy of crude methanol extract of Beriain MJ, Bas P, Purroy A, Treacher T, Ledin I, Morand-Fehr P Donnetia Tripetala Fruits with its various chemical fractions in mice (2000). Effect of animal and nutritional factors and nutrition on lamb experimentally infected with Heligmosomoides. Master’s Degree meat quality. InProceedings of the 8th Seminar of the Sub-Network Dissertation. Department of Veterinary Pharmacology and on Nutrition of the FAO-CIHEAM Inter-Regional Cooperative Toxicology, Ahmadu Bello University, Zaria, Nigeria. 124 p. Research and Development Network on Sheep and Goats, pp. 3-5. Knox MR, Torres-Acosta JFJ, Aguilar Caballero AJ (2006). Exploiting Bermingham EN, Nabb Mc, Sutherland WC, Sinclair IABR, Treloar B P, the effect of dietary supplementation of small ruminants on resilience Roy NC (2007). Adult Trichostrongylus colubriformis infection did not and resistance against gastrointestinal nematodes. Vet. Parasitolol. affect protein synthesis rate in whole-body, intestinal, hepatic and 139:385-393. skeletal muscle tissues of lambs fed fresh Lucerne (Medicago sativa) Lopez V, Lindsay RC (1993a). Metabolic conjugates as precursors for Can. J. Anim. Sci. 87(3):315-325. characterizing flavor compounds in ruminant milks. J. Agric. Food Biu AA, Olatunde KT, Konto M, Paul BT, Ya’uba AM (2012b). Chem. 41:446-454. Prevalence and quantitative faecal analysis of equine gastrointestinal Mbafor Fidelia Lem V, Khan Payne J, Wabo Poné, Mbogning Tayo nematodosis in Maiduguri, Nigeria. CENRESIN. J. Med. Appl. Biosci. Gertrude J, Tchoumboue (2014). In vivo Anthelmintic Activity of 4:1-6. Terminalia glaucescens (Combretaceae) Extracts against Borokini TI, Omotayo FO (2012). Comparative phytochemical analysis Gastrointestinal Nematodes of Sheep. British J. Pharm. Res. 4:18. of selected medicinal plants in Nigeria Int. J. Adv. Chem. Miller M. (2007). Dark, firm and dry beef. National Cattlemen's Beef Res.1(1):011- 018. Association, on behalf of The Beef Checkoff Program. U.S.A.: Cheng Q, Sun DW (2008). Factors affecting the water holding capacity Centennial, CO. of red meat products: A review of recent research advances. Critical Mohammed Konto , Yusuf Abba, Nur Syairah Binti Ramli, Murugaiyah Rev. Food Sci. Nutr. 48(2):137-59. Marimuthu, Mohammed Ariff Omar, Faez Firdaus Jesse Abdullah, Cvrtila Fleck Z, Kozačinski B, Njari D, Marenčić G, Mršić K, Spiranec Muhammad Abubakar Sadiq, Abdulnasir Tijjani, Eric Lim Teik D, Špoljarić MJ, Čop M, Ţivković M, Popović (2015). Chung, Mohammed Azmi, Mohammed Lila (2016). The use of Technological properties and chemical composition of the meat of FAMACHA in estimation of gastrointestinal nematodes and total sheep Technological properties and chemical composition of the worm burden in Damara and Barbados Blackbelly cross sheep. Trop.

Germaine et al. 139

Anim. Health Prod. 48:1013-1020. Agric. Sci. 110(3):531-541. Moyo B, Masika PJ, Muchenje V (2014). Effect of feeding Moringa Thienpont D, Rochette F, Vanparijs OFJ (1986). Diagnosing (Moringa oleifera) leaf meal on the physico-chemical characteristics Helminthiasis by Coprological Examination. Janssen Reseach and sensory properties of goat meat. S/Afr. J. Anim. Sci. 44:64-70. Foundation, Beerse Belgium. 205p. Mushi DE, Mtenga LA, Eik LO (2007). Some factors affecting the quality Tsiboukis D, Sazakli E, Jelastopulu E, Leotsinidis M (2013). of meat from ruminants and their relevance to the Tanzanian meat Anthelmintics residues in raw milk. Assessing intake by a children industry - review. Tanzania J. Agric. Sci. 8:173-192. population. Polish J. Vet. Sci.16:85-91. Ndamukong KJN, Sewell MM, Asanji MF (1989). Management and Valieva Z, Sarsembaeva N, Valdovska A, Ussenbayev AE (2014). Productivity of small ruminants in the North West Province of Impact of Echinococcosis on Quality of Sheep Meat in the South Cameroon. Trop. Anim. Health Prod. (21):109-119. Eastern Kazakhstan. Asian-Austral. J. Anim. Sci. 27(3):391-397. Oloyede OI, Aluko OM (2012 ). Determination of Antioxidant Potential of Vargas-Magaña JJ, Torres-Acosta JF, Aguilar-Caballero AJ, Sandoval- Momordica foetida Leaf Extract on Tissue Homogenate. Sci. J. Med. Castro CA, Hoste H, Chan Pérez JA. (2014). Anthelmintic activity of Clin. Trials, 225. acetone-water extracts against Haemonchus contortus eggs: Olukayode MO, Adebola OO (2008). Antibacterial activity of crude and interactions between tannins and other plant secondary compounds. Fractions of M. feotida leaf extracts. Int. J. Biomed. Pharm. Sci.; Vet. Parasitol. 206(3-4):3227. 2(2):75-78. Vilegs GH, DeMarchi E , Lancas FM (1997). Extraction of low polarity Page JK, Wulf DM, Schwotzer TR (2001). A survey of beef muscle compounds (with emphasis on coumarin and kaurenoic acid) from colour and pH. J. Anim. Sci. 79:678-687. Mikania glomerata (guaco) leaves. Phytochem. Anal. 8:266-270. Pauwels JM E, van Ranst M, Verloo A, MvondoZe (1992). Manuel de Williams Andrew R, Palmer Dieter G (2012). Interactions between Laboratoire de pédologie. Publications Agricoles N° 28. Bruxelles: gastrointestinal nematode parasites and diarrhoea in sheep: AGCD Pathogenesis and control. Vet. J. 192:279-285. Pavan Kumar1 PV, Naga S, Subrahmanyam S (2013). Phytochemical Williams P, Droulez V, Levy G (2006). Nutrient composition of analysis, in-vitro screening for antimicrobial and anthelmintic activity Australian red meat (2002). Gross composition data. Food Aust. of combined hydroalcoholic seed extracts of four selected folklore 58:173-181. indian medicinal. Plants Scholars Res. Library. 5(1):168-176. Young OA, Reid D H, Scales GH (1993) Effect of breed and ultimate pH Phillips K, Wheeler K (2008). Investigation of the variation in lamb meat on the odour and flavour of sheep meat. New Zealand J. Agric. Res. quality on three winter finishing systems. ADAS Lincoln, Ceres 36(3)363-370. House, 2 Searby Road, Lincoln LN2 4DW. Final Report Paper, Zhong RZ, Li HY, Fang Y, Sun HX, Zhou DW (2015). Effects of dietary ADDAS, XLB1424 supplementation with green tea polyphenols on digestion and meat Piasentier Em, Valusso R, Volpelli LA, Failla S, (2003). Meat quality of quality in lambs infected with Haemonchus contortus. Meat Sci. Simmental young bulls as affected by the gene frequency of 105:1-7. Montbeliarde origin. In Proceedings 49th international congress of meat science and technology. August 31st to September 5th, Campinas, Sao Paulo, Brazil. pp. 187-188. Qwele K, Hugo A, Oyedemi SO, Moyo B, Masika PJ, Muchenje V (2013). Chemical composition, fatty acid content and antioxidant potential of meat from goats supplemented with Moringa (Moringa oleifera) leaves, sunflower cake and grass hay. Meat Sci. 93:455- 462. Rajib SMFR, Dey AR, Begum N, Momin MA, Talukder MH (2014).Prevalence and in vitro culture of Trichostrongylus spp. In goat at Trishal, Mymensingh, Bangladesh. J. Adv. Parasitol. 1(4):44 - 48. Rinaldi M, Leentje D, Prisca RH, Robert WL, Edwin C, Bruno G, Jozef V, Wim Van DB, Peter G (2011). Infection with the gastrointestinal nematode Ostertagia ostertagi in cattle affects mucus biosynthesis in the abomasum. Veterinary Research 42:61. https://doi.org/10.1186/1297-9716-42-61 Roy EA, Hoste H, Beveridge I (2004). The effects of concurrent experimental infections of sheep with Trichostrongylus colubriformis andT. vitrinus on nematode distributions, numbers and on pathological changes. Parasit. 3:293-300. Samad MA (2008). Animal Husbandry and Veterinary Science. LEP Publication.No.11, BAU campus, Mymensingh, Bangladesh. 244 – 245. Sinclair A, Mann N, O'Connell S, (1999). The nutrient composition of Australian beef and lamb. Melbourne: RMIT. Smallholders in Southeast Asia. In: Sani, R.A.; Gry, D.G. and Baker, R.L.: Worm control for small ruminants in tropical Asia (Canberra, Australia). Aciar Monograph. 113:51-61. Stear MJ, Bishop SC, Henderson NG, Scott I (2003). A key mechanism of pathogenesis in sheepinfected with the nematode Teladorsagia circumcincta. Anim. Health Res. Rev. 4(1):45-52. Steel JW, Jones WO, Symons LEA (1982). Effects of a concurrent infection of Trichostrongylus colubriformis on the productivity and physiological and metabolic responses of lambs infected with Ostertagia circumcincta. Austral. J. Agric. Res. 33(1):131-140. Sykes AR, Poppi DP, Elliot DC (1988). Effect of concurrent infection with Ostertagia circumcincta and Trichostrongylus colubriformis on the performance of growing lambs consuming fresh herbage. J.

Vol. 12(6) pp. 140-150, June 2018 DOI: 10.5897/AJFS2018.1709 Article Number: A41B40957077 ISSN 1996-0794 Copyright © 2018 Author(s) retain the copyright of this article African Journal of Food Science http://www.academicjournals.org/AJFS

Full Length Research Paper

Comparison of traditional butter preservation techniques using microbial and organoleptic properties, West Shewa, Ethiopia

Alganesh Tola1*, Debela Bayu3, Lemma Fita4, Bilatu Agza2 and Sara Birkie2

1Holeta Agricultural Research Center, Ethiopian Institute of Agricultural Research, P. O. Box 31, Holeta, Ethiopia. 2Agricultural Quality Research laboratories, Ethiopian Institute of Agricultural Research, P. O. Box 2003, Addis Ababa, Ethiopia. 3Department of Animal Sciences, Faculty of Agriculture, Asosa University, P. O. Box, Asosa, Ethiopia. 4Department of Animal Sciences, Faculty of Agriculture and Veterinary Medicine, Ambo University,P. O. Box 19, Ambo, Ethiopia.

Received 16 March, 2018; Accepted 26 April, 2018

The study was conducted in Dire Inchini and Ambo districts of West Shewa, Oromia, Ethiopia to assess traditional butter preservation methods and compare their efficiency. Semi-structured questionnaire was prepared, pretested and used to interview 120 women respondents having experience in butter making. Butter preservation methods identified were ghee making (100%), spicing (98.33%), melted butter (29.17%) and salting (11.67%). Commonly used spices were Trachyspermum ammi, Nigella sativa, Aframomum angusti-folium, Trigonella foenum, Zingiber officinale and Allium sativum. Based on the survey, 7 kg of fresh butter samples were purchased from open market in the two districts and taken to Holeta Dairy laboratory and randomly allocated to each treatment namely traditional ghee, untreated, salted, spiced, melted, frozen (-20°C) and refrigerated butter (4°C). Microbial and organoleptic qualities of butter were analyzed at one month interval for 3 consecutive months. Microbial qualities of the samples were substandard; but, traditional ghee and salting were more efficient. Optimization of utilization of spices and comprehensive evaluation including oxidative deterioration is vital.

Key words: Traditional, butter, preservation, microbial, organoleptic quality.

INTRODUCTION

Like most sub Saharan Africa countries, Ethiopia is Ethiopia is processed into milk products at household unable to meet the increasing demand for dairy products level using traditional technologies (Muriuki and Thorpe, for its increasing population (Azage et al., 2000; Tsehay, 2001). In rural areas, 40% of milk produced is 2001). Smallholder farmers and pastoralists produce and spontaneously fermented for three to four days without supply 98% of the total annual milk production (Yonad, addition of specific starter culture and is churned to make 2009). The majority of milk produced in rural areas of butter, butter milk and whey as a byproduct. Rural

*Corresponding author. E-mail: [email protected].

Author(s) agree that this article remain permanently open access under the terms of the Creative Commons Attribution License 4.0 International License Tola et al. 141

producers are forced to produce butter due to limited Sampling techniques market outlet, shorter shelf life of milk, lower price for whole milk, ease of handling of butter and for product The survey was conducted in three major ecologies of the districts as follows: Ya’i cabo (high altitude), Gosu qora (mid altitude), diversification (Ayantu, 2006; Kassahun, 2008). Sankele and Farisi ‘(low land) and, Waldo hindhe (high land), Nano Traditional butter ferments slowly at ambient temperature, Jidu (mid altitude) and Toke Abuye (low land) from Ambo and Dire offering rural consumers a relatively shelf stable dairy Inchini district, respectively. product (LMP, 2007). According to Getachew (2003), of the total butter production, 80% is used as food ingredient and the remaining is used for hair dressing and Data collection other purposes. The same study revealed that 70% of Survey butter produced in the country is used in rural areas; while 30% is channeled to the Addis Ababa market. One hundred twenty respondents who own at least one lactating Therefore, butter contributes much to the dietary cow and having experience in traditional butter preservation requirements of the society and saves milk from spoilage techniques were purposively selected. Semi structured and diversifies its use (FAO, 1990). Workneh and Ulfina questionnaire was prepared, pretested and used for individual interview by trained enumerators. The questionnaire focused on (2011) observed the significant importance of butter in butter preservatives and preservation methods. The collected data Ambo area of West Shoa Zone and indicated that 75% of was entered into SPSS computer program. dairy producers in Ambo area process milk into butter. However, in Ethiopia, the climate is hot and humid; leading dairy products to spoil easily during storage Laboratory analysis unless cooled or treated with preservatives. Moreover, Collection of butter samples commercial preservatives are not readily available in rural areas. Cooling systems are also not feasible because of Seven kilograms of butter was bought from open markets of the two lack of infrastructure (O’Mahoney and Peters, 1987). In districts to assess the microbial and organoleptic qualities. The rural areas of Ethiopia, producers use different traditional purchased butter samples were kept in an icebox and transported preservation methods to increase shelf life of butter to Holeta dairy laboratory within 4 h of collection. The samples were (Lemma, 2004; Mekdes, 2008). The local preservatives thoroughly mixed to form composite sample which was divided into seven equal weights each treatment receiving one kilogram of are used as a principle of acidification and moisture butter. The samples were randomly allocated to seven treatments reduction and can make butter of good storage stability namely traditional ghee, spiced, salted, melted, untreated, frozen (- (O'Mahony and Peters, 1987). However, traditional butter 20°C) and butter stored at 4°C. All analysis was done using preservation techniques and their efficiency in these duplicate samples. districts were not studied. Therefore, the study was initiated to assess traditional butter preservation Preparation of treatments techniques and to compare their efficiency through evaluation of microbial and organoleptic properties. The samples were tested for their microbial and organoleptic qualities at 0, 1, 2 and 3 months of preservation. The treatments were prepared as follows. MATERIALS AND METHODS

Description of the study areas Traditional ghee/‘Nitir Kibe’ The study was conducted in Ambo and Dire Inchini districts of West Shoa Zone of Oromia, Ethiopia which is located at 105 km to the One kilogram butter was placed in a sauce pan and melted over a west of Addis Ababa, Ethiopia. slow heating stove. White cumin, fenugreek, korerima, ginger, garlic, turmeric, black cumin and other herbs such as basil and other herbs of desirable aroma such as rue (Ruta graucolence), Ambo district basil (Ocimum spp.) and ‘Kussayyee’(Ocimum hardiense) aroma were added to the boiling butter fat based on the respondents’ It is located between 80 47’N -90 21’N and 370 32’E and located at experience. The melting butter fat and spices were stirred while 115 km to the West of Addis Ababa (ADADO unpublished data, boiling until foaming had stopped. Finally, the sauce pan was 2014). It has a mean annual temperature and rain fall of 23 to 25°C removed from the stove and left aside until it was settled. The butter and 1300 to 1700 mm, respectively. The altitude ranges between fat was filtered through metal sieve into high density poly ethylene 500 and 3,200 m above sea level (ADADO Unpublished Data, bucket and kept at room temperature for a period of 3 months. 2014).

Spiced butter Dire Inchini district One kilogram of butter was removed from composite butter sample It is located at 162 km to the West of Addis Ababa and 50 km from and thoroughly mixed with about 45 g of fenugreek and black cumin Ambo (Tamrat, 2007). The average annual rainfall ranges between powders, respectively. Based on the recommendation of the 1000 and 1400 mm. Average minimum and maximum temperatures respondents, spiced butter sample was placed in high density poly are 8.8 and 21.6°C, respectively (Tamrat, 2007). ethylene bucket and kept at room temperature for 3 months. 142 Afr. J. Food Sci.

Salted butter plating agar media of 3 to 4 ml to inhibit surface colony formation. The medium were allowed to solidify. The plates were inverted and One kilogram butter was thoroughly mixed with 30 g of NaCl and incubated at 32 ±1°C for 24±2 h. Counts were made manually. kept in high density poly ethylene bucket and was placed at room Finally, the plate counts were calculated as N, the number of colony temperature for 3 months. forming units of coliforms per milliliter of milk sample using the formula N=∑c/(n1+n2)d where ∑ c= Sum of all colonies on all plates counted were conducted according to the standard procedures of Melted butter/‘Nigur kibe’ FAO (1997), Michael and Jospeh (2004) and FSSAI (2012).

To make melted butter one kilogram of butter was placed in sauce Lactic acid bacteria counts: 0.1 ml of appropriate decimal pan and kept on a slow heating stove and stirred until the butter dilutions of butter sample was poured on Petri dishes in duplicates was completely melted. The melted butter was removed from the and mixed with MRS agar media. Then, after incubating the plates heat source and kept in cool place to settle down and left there for anaerobically at 30°C for 48 h, lactic acid bacteria were counted overnight until it has completely solidified. Then, impurities that (FAO, 1997; Michael and Joseph, 2004; FSSAI, 2012). were settled at the bottom of the sauce pan were decanted off by opening the solidified butter in sauce pan from one side. Then the Yeast and mold counts: Potato dextrose agar (PDA) media was sample was placed in high density poly ethylene bucket and kept at autoclaved for 15 min at 121°C and tempered in water bath room temperature for 3 months. adjusted at 45°C. Appropriate decimal dilutions of butter sample (0.1 ml) were poured into a 15 × 90 ml Petri dishes and mixed with 20 ml of PDA containing antibiotic; chloramphenicol solution. Then, Untreated butter after incubation at 25°C for 5 days, yeast and mould were counted with plates containing 10 to 150 colonies (FAO, 1997; Michael, One kilogram of fresh butter sample was kept in high density poly 2004; FSSAI, 2012). ethylene bucket and was placed at room temperature for 3 months. Enterobacteriaceae count: One milliliter of homogenized melted butter sample was added into 9 ml peptone water to yield a dilution Frozen butter of 1:10 cfu/ml. Violet red bile glucose agar (VRBGA) medium was used to enumerate Entrobacteriaceae. The mixture was then One kilogram of butter sample was kept in high density poly overlaid with the same plating agar media of 3 to 4 ml. Plates were ethylene bucket and placed in deep freeze at -20°C for 3 months. aerobically incubated for 24 h at 37°C and inspected for purple-red colonies surrounded by a purplish circle of light or halo color. The

plate counts were calculated by multiplying the count on the dishes Refrigerated butter n by 10 , where n stands for the number of consecutive dilutions of

the original sample (ILSI, 2011, FSSAI, 2012). One kilogram of composite butter sample was kept in high density poly ethylene bucket and placed in refrigerator at 4°C for 3 months. Organoleptic quality of butter: Organoleptic quality were Each time at 0, 1, 2 and 3 months of preservation, required evaluated by 10 semi-trained sensory panelists using 5 point amount of sample was removed for analysis of aerobic mesophilic hedonic rating scale where 1=dislike extremely, 2= dislike slightly, bacteria, total coliforms, total lactic acid bacteria, 3= neither like nor dislike, 4= like moderately and 5 = like extremely. Enterobacteriaceae and yeast and mold counts and color, texture, The samples were coded for identification purpose. odor and overall acceptability. The analysis of each treatment was performed in duplicates. Data analysis

Microbial analysis Survey and organoleptic quality data were analyzed using descriptive statistics using SPSS (2011). Microbiological counts Aerobic mesophilic bacterial count: The butter samples were were transformed to log10 cfu/g and analyzed using the General homogenized and aseptically transferred to stomacher bag and Linear Model of SAS version 9.1 (SAS, 2009). Least significant ° held for melting on a water bath adjusted at 46 C. Then, the difference was used to test the differences between treatment samples were immediately serially diluted by adding 1 ml of butter means and time of preservation. into 9 ml of peptone water. One milliliter of the sample from a chosen dilution was placed on the Petri dish using pour plating technique. Then, plate count agar media of 15 to 20 ml was poured RESULTS AND DISCUSSION onto the petri-dish and thoroughly mixed with the sample and allowed to solidify for 15 min and incubated for 48+2 h at 35°C. Traditional butter preservation techniques in the Finally, the colonies were manually counted. The plate counts were n study areas calculated by multiplying the count on the dish by 10 , in which n stands for the number of consecutive dilutions of the original Traditional methods of butter preservation in Dire Inchini sample (FAO, 1997; Michael and Joseph, 2004; FSSAI, 2012). and Ambo districts were presented in Table 1. In the

Total coli forms count: Samples were decimally diluted and plated study areas, 100, 98.33, 29.17 and 11.67% of interviewed with violet red bile agar media /VRBA into Petri dishes for smallholder producers commonly preserve butter in the enumeration of total coliforms bacteria as coliforms colony forming form of traditional ghee, spiced, melted and salted butter, units per milliliter. Plates were incubated at t 32±1°C for 24±2 h. respectively. This finding is in agreement with previous One milliliter of melted sample was serially diluted using peptone reports in Arsi Negele, Oromia, Walayita, Southern water and transferred into sterile Petri-dishes. 10 to 15 ml of violet red bile agar media tempered to a temperature of 44 to 46°C was Ethiopia, North Western Ethiopia and East Wollega, added to the milk sample and thoroughly mixed and allowed to Ethiopia by Lemma et al. (2004), Mekdes (2008), Eyassu solidify for 5 to 10 min. The mixture was then overlaid with the same (2014) and Alganesh and Yetenayet (2016), respectively. Tola et al. 143

Table 1. Traditional methods of butter preservation in the study areas.

Preservation method Dire Inchini (N=60) Ambo (N=60) Overall mean (N=120) Traditional ghee N % N % N % Mixing with spices 60 100 60 100 120 100 Melting 58 96.67 60 100 118 98.33 Salting 20 33.33 15 25 35 29.17

N = Number of respondents.

Table 2. Spices used for butter preservation techniques in the study areas.

Local name Common name Scientific name N (%) Habasuuda adii White cumin Trachyspermum ammi 120 (100) Ogiyoo Kororima Aframomum angusti-folium 98(81.67) Abishii Fenugreek Trigonella foenum 85(70.83) Jinjibila Ginger Zingiber officinale 71(59.17) Qullubbii adii Garlic Allium sativum 61(50.83) Irdii Turmeric Curcuma domestica 3(2.5) Qarafaa Cinnamomum verum 3(2.5) Dimbilaala Basil Cordiandrum sativum 1(0.83) Qullubbii diimaa Onion Allium cepa 3(2.5) Qundeberbere Black peper Piper nigrum 2(1.67) Habasuuda gurraacha Black cumin Nigella sativa 104(86.67)

N= number of respondents.

In the study areas in wet season and during Ethiopian treatments and preservation time is presented in Table 3. Orthodox fasting time, demand for butter is low and The aerobic mesophilic bacterial counts of butter during such occasions, surplus butter is preserved using preserved using traditional ghee, salted and spiced butter traditional preservation techniques. In the present study, at 0, ends of 1, 2 and 3 months of preservation did not spiced, melted and salted butter are used as raw show significant difference at P<0.05. While aerobic materials for traditional ghee making and they are mesophilic bacterial counts of melted, untreated, frozen optional butter preservation techniques. and butter stored in refrigerator at 4°C showed significant difference among treatments at 0, ends of 1, 2 and 3

Spices used for ghee making and spicing butter months at P<0.05 level of significance. The total aerobic (spiced butter) mesophilic bacterial counts for traditional ghee, salting, spicing, melted, untreated, frozen and refrigerated butter The result for the spices used for spiced butter and (40c) showed significant difference among the treatment traditional ghee making in the study areas were indicated means at P<0.05 level of significance. At initial in Table 2. For traditional ghee making smallholder preservation time, greater counts of aerobic mesophilic producers in the study areas mainly use Trachyspermum bacterial counts was observed in salted (9.58 log cfu/g) ammi, Aframomum angusti-folium, Nigella sativa, and melted butter (9.65 log cfu/g), which were Trigonella foenum, Zingiber officinale, Allium sativum and significantly different (P<0.05) from other treatments. This Curcuma domestica is used for coloring purpose. While might be due to the poor hygiene of the salt used in for making spiced butter, they use Trigonella foenum and preserving the butter. In the case of melted butter, the Nigella sativa. Similar study by Joe et al. (2009) stated increase in the aerobic mesophilic bacterial count might that spices are used to enhance aroma, flavor and for probably be as a result of post melting contamination. At preservation of food substances. initial preservation period, the mean aerobic mesophilic bacteria in untreated butter were 8.71 log cfu/g of butter samples. The current result is far beyond the maximum Microbial properties of butter preserved using tolerable limit of 6 log cfu/g of aerobic mesophilic bacteria traditional preservation methods counts set by Standards Authority of Ethiopia (QSAE, Aerobic mesophilic bacterial count 2009). The sensory attributes used to evaluate butter samples in this experiment were odor, texture, color and The mean total bacterial count (log cfu/g) for the overall acceptability. So there was no risk on the health 144 Afr. J. Food Sci.

Table 3. Mean total aerobic mesophilic bacterial counts (log cfu/g) of traditionally preserved butter.

Time Traditional Spiced Untreated Refrigerated Maxlimit set Variable Salted butter Melted butter Frozen butter (month) ghee butter butter butter by ESA 0 9.48±0.08aB 9.39±0.09aB 9.58±0.05aB 9.65±.02aB 8.71±.03bC 8.71±.02aCD 8.71±0.02bD Aerobic 1 9.62±0.03aA 9.37±.06aB 9.62±0.04aB 9.72±0.05aB 9.61±0.03aB 9.59±0.09aC 9.59±0.09aB mesophilic aA aA aA bC aB aB aB 6 bacterial 2 9.64±0.04 9.66±0.06 9.84±0.04 9.23±0.06 9.68±0.04 9.61±.04 9.63±0.05 3 9.73±0.06aA 9.78±0.03aC 9.89±0.06aA 9.93±0.03aA 9.98±0.04aA 9.24±.04bD 9.97±0.03aB

Means followed by similar lower case letters in a column are not significantly different (P<0.05), Means followed by similar upper case letters in a row are not significantly different (P<0.05). ESA: Ethiopian Standard Authority.

Table 4. Mean total lactic acid bacteria counts (log cfu/g) of traditionally preserved butter.

Time Traditional Salted Spiced Melted Untreated Frozen Refrigerated Variable (month) ghee butter butter butter butter butter butter 0 6.65±0.09aA 6.55±0.05aA 6.72±0.06aA 6.80±0.28aA 6.77±0.03aA 6.77±0.08aA 6.77±0.07aA bC abC bD bC aB aB aB Total lactic acid 1 6.06±0.06 6.31±0.06 6.09±0.13 6.40±0.05 6.65±0.09 6.23±0.04 6.59±0.04 bacteria 2 5.56±0.06cC 5.19±0.09cC 5.72±0.05cC 5.26±0.07cC 6.60±0.07aB 6.28±0.08aB 6.12±0.10aC 3 5.09±0.11cD 5.01±0.17cD 5.53±0.04bD 5.01±0.14cD 6.32±0.07aC 6.04±0.10aC 6.06±0.10aD

Means followed by similar lower case letters in a column are not significantly different (P<0.05), Means followed by similar upper case letters in a row are not significantly different (P<0.05)

and safety of the panelists since they evaluated of spices, heat treatment and low temperatures in described in Table 4. There were significant the butter by smelling to check for the odor, spiced butter, traditional ghee, melted butter, differences (P<0.05) between total lactic acid checked the texture using smoothness, solidity butter kept at 4 and -20°C, respectively on bacterial counts in traditional ghee, salted, spiced and appropriate degree of firmness by their hands bacterial growth and survival. Similar studied by and melted butter at 0, ends of 1, 2 and 3 months and visual observation of the color of the butter Kilcast and Subramaniam (2000) confirmed that of preservation. While the total lactic acid bacterial samples. The current result is also similar to a shelf life of products can be extended by the use counts of butter preserved using untreated, frozen report from Wolayita in Southern Ethiopia of 8.10 of processing treatments such as heat and (-20°C) and refrigerated (4°C) butter showed no log cfu/g of butter samples of total bacterial count radiation which kills the microorganisms or control significant difference at P<0.05 among the means (Mekdes, 2008). After one month of preservation, of microbial growth by chilling, freezing, reducing at 0, 1, 2 and 3 months of preservation, aerobic mesophilic bacterial counts of 9.37 log the water content and addition of preservatives. respectively. Total lactic acid bacterial counts at 0 cfu/g was observed and this was significantly and 1 month of preservation showed no significant different (P<0.05) from the other treatments. (P<0.05) difference for traditional ghee, salted, Relatively lower mean aerobic mesophilic Lactic acid bacterial count spiced, melted, untreated, frozen and refrigerated bacterial counts were observed in frozen butter, (4°C) butter samples. While, the total lactic acid traditional ghee and spiced butter. This might be The mean total lactic acid bacteria counts (log bacterial counts at the end of the second month of due to the antagonistic effects of active ingredients cfu/g) of treatments and preservation time are preservation did not show significant (P<0.05) Tola et al. 145

Table 5. Mean yeast and mold counts (log cfu/g) of traditionally preserved butter.

Time Traditional Salted Spiced Melted Untreated Frozen Refrigerated Max. limit set Variable (month) ghee butter butter butter butter butter butter by ESA 0 5.70±0.08aB 6.54±0.03bC 5.74±0.05aC 5.56±0.03dD 6.70±.05bD 6.70±0.04aA 6.70±0.05aB aB aB aA cC cC bB aB Yeast and 1 5.70±0.04 6.67±.03 6.36±0.05 5.78±0.09 6.74±0.03 6.46±0.05 6.72±0.06 1 molds 2 6.16±0.08aA 6.69±.04aB 6.28±0.08aB 6.28±.07bB 6.78±.03aB 6.14±0.10bC 6.76±0.05aA 3 6.19±0.08aA 6.78±0.05aA 6.37±0.06aA 6.71±.07aA 6.84±.05aA 6.42±0.14bC 6.79±0.04aA

Means followed by similar lower case letters in a column are not significantly different from each other (P<0.05), Means followed by similar upper case letters in a row are not significantly different from each other (P<0.05). ESA: Ethiopian Standard Authority

difference among traditional ghee, salted, spiced, treatments. This might be due to heat treatment attributable to the effect of heat treatment; melted and refrigerated butter (4°C) except for and antimicrobial effects of spices used in melted antimicrobial properties of spices used to treat untreated and frozen butter (-20°C). Whereas, at butter, traditional ghee and spiced butter, butter samples and reduced water activity in the end of the third month of preservation, the respectively. salted butter. The temperature range for yeast mean total lactic acid bacterial counts in traditional and mold growth is 0 to 47°C, out of which, the ghee, salted , spiced, melted and refrigerated growth of yeast and mold can be hampered. This butter samples did not show significant (P<0.05) Yeast and mold counts is in agreement with Seriler (2003) who revealed difference among their mean counts except for the possibility of reducing mold growth on the untreated and frozen butter which did not The result for the mean yeast and mold counts surface of butter by salting. The mean yeast and significantly (P<0.05) differ from each other. The (log cfu/g) of treatments and preservation time is mold count of untreated butter sample at initial mean total lactic acid bacterial counts of the indicated in Table 5. The mean yeast and mold time of preservation was 6.70 log cfu/g. The treatments at initial time of preservation ranged counts during initial 0, 1, 2 and 3 months of current finding is beyond the maximum tolerable from 6.55 to 6.80 log cfu/g. This could be due to preservation for traditional ghee, spiced and butter limit of 1 log cfu/g of yeast and mold count of prior fermentation of composite sample as local refrigerated at 4°C did not significantly (P<0.05) butter sample recommended by the Ethiopian butter is usually made of spontaneously fermented differ from each other. The mean yeast and mold Standards Authority (QSAE, 2009). The present whole milk. At initial time of preservation, the counts (log cfu/g) of salted, spiced, melted, result is also higher than the mean yeast and mean lactic acid bacterial count in traditional ghee untreated and frozen butter samples significantly mould count of 5.58 log cfu/g of butter samples was 6.06 log cfu/g and it significantly differed at differed (P<0.05) from each other. The mean reported in Wolayita Southern Ethiopia (Mekdes, P<0.05 from other treatments; except for salted yeast and mold counts (log cfu/g) of traditional 2008). Increasing trends of yeast and mold counts butter which was 6.09 log cfu/g of butter samples. ghee, salted, spiced, melted, untreated, frozen have been observed from one month of The current result is also similar with the finding of and refrigerated butter samples did not show preservation period to the end in untreated, Mekdes (2008) who reported mean lactic acid significant difference (P<0.05) between the refrigerated (at 4°C), spiced and melted butter and bacterial count of 7.51 log cfu/g for butter sample treatment means. Mean yeast and mold counts of these results were significantly different (P<0.05) collected from Wolayita in Southern region. From butter at initial preservation time for traditional from other treatments. In the case of spiced butter the second to third months of preservation time, ghee (5.70 log cfu/g), salted butter (5.74 log cfu/g) poor hygiene of spices purchased from open mean lactic acid bacterial count of spiced, melted and melted butter (5.56 log cfu/g) significantly market might have contributed to the high rate of butter and traditional ghee were relatively lower differed (P<0.05) from spiced, untreated, frozen (- contamination. In untreated and refrigerated butter and significantly differed (P<0.05) from other 20°C) and refrigerated (4°C) butter. This might be samples water activity might have been high and 146 Afr. J. Food Sci.

Table 6. Mean total coliform counts (log cfu/g) of traditionally preserved butter.

Time Traditional Salted Spiced Melted Untreated Frozen Refrigerated Variable ESA (month) ghee butter butter butter butter butter butter 0 5.70±0.08aA 6.54±0.03aB 5.74±0.05aC 5.56±0.03aD 6.70±0.05aB 6.70±0.04aA 6.70±0.05aA aA aB aB aC bC aA aA Total 1 5.70±0.04 6.67±0.03 6.36±0.05 5.78±0.09 6.74±0.03 6.46±0.05 6.72±0.06 Absent coliform 2 6.16±0.08aA 6.69±0.04aB 6.28±0.08aB 6.28±0.07aB 6.78±0.03aB 6.14±0.10aA 6.76±0.05aA 3 6.37±0.08aA 6.96±0.11aA 6.54±0.05aC 6.86±0.09aB 6.98±0.09aA 6.34±0.07aA 6.62±.05aA

Means followed by similar letters in a column are not significantly different from each other (P<0.05); means followed by similar bold letters in a row are not significantly different from each other (P<0.05). ESA: Ethiopian standard Authority.

have created favorable environment for the growth (P<0.05) differ from each other. At the end of treatments and preservation time is presented in of yeast and molds. second month of preservation, the mean total Table 7. There were no significant difference coliforms counts of traditional ghee, spiced, (P<0.05) between mean Entrobacteriaceae counts melted, frozen and refrigerated butter samples did of traditional ghee, salted, spiced, melted, frozen Total coliform counts not significantly (P<0.05) differ from each other and refrigerated butter except for untreated butter except for salted and untreated butter. At the end at 0, 1, 2 and 3 months of preservation, Mean total coliforms count (log cfu/g) for the of the third month of preservation, the mean total respectively. The mean Entrobacteriaceae counts treatments and time of preservation is presented coliforms counts of traditional ghee, spiced, of butter samples for traditional ghee, spiced, in Table 6. There were no significant difference melted, frozen, refrigerated, salted and untreated melted and frozen butter at 0 month of (P<0.05) between the mean total coliforms butter samples did not significantly (P<0.05) differ preservation did not significantly (P<0.05) differ bacterial counts of butter preserved using from each other. Mean total coliform count of from each other. While the mean traditional ghee, salted, frozen and refrigerated untreated butter at initial preservation period was Entrobacteriaceae counts of butter samples for butter at 0, 1, 2 and 3 months of preservation, 5.62 log cfu/g of butter sample. The current result salted, untreated and refrigerated butter at initial while, the total coliforms bacterial counts of is by far beyond the mean total coliform count of 2 preservation time significantly (P<0.05) differed spiced, melted and untreated butter showed log cfu/g of butter samples collected from from other treatments but the mean counts for significant (P<0.05) difference among treatments Wolayita zone (Mekdes, 2008). From two months untreated and refrigerated butter did not at 0, 1, 2 and 3 months of preservation, of preservation period and onwards, relatively significantly differ from each other. The mean respectively. Mean total coliform counts at initial decreasing trends of total coliform count were Entrobacteriaceae counts of samples for time of preservation for traditional ghee, spiced, observed in traditional ghee, frozen and salted traditional ghee, spiced, melted and frozen butter melted, untreated, frozen and refrigerated butter butter compared to other treatments. This might at the end of one month did not significantly samples did not differ (P<0.05) significantly except be associated with the inhibitory effects of heat (P<0.05) differ from each other. While, the mean for salted butter. The mean total coliforms counts treatment, low storage temperature and addition Entrobacteriaceae counts of samples for salted, at the end of one month of preservation for of salt in ghee, butter kept at-20°C and in salted untreated and refrigerated butter at initial traditional ghee, spiced, melted, frozen and butter, respectively. preservation time significantly (P<0.05) differed refrigerated butter samples were not significantly from other treatments. But the mean counts for different at P<0.05 except for salted and untreated untreated and refrigerated butter did not butter. While the mean total coliform counts at the Enterobacteriaceae count significantly differ from each other. The mean end of one month of preservation for salted and Entrobacteriaceae counts of butter samples at the untreated butter samples did not significantly Mean Entrobacteriaceae count (log cfu/g) for the end of second months of preservation for Tola et al. 147

Table 7. Mean Entrobacteriaceae counts (log cfu/g) of traditionally preserved butter.

Time Traditional Salted Spiced Melted Untreated Frozen Refrigerated Variable EUS limit (month) ghee butter butter butter butter butter butter 0 4.26±0.04aA 6.70±0.07aB 5.83±0.06aA 6.27±0.03aA 5.07±0.08cC 5.07±0.33aA 5.07±0.31aC 1 5.79±0.08aB 6.84±0.03aA 5.94±0.06aB 6.42±0.03aB 5.64±.04aC 5.57±0.04aB 5.67±0.05aC Enterobacteriaceae 3 2 5.84±0.06aA 6.52±0.03aB 5.97±0.06aA 6.57±0.07aB 6.23±0.09aB 5.69±0.04aB 5.96±0.11aA 3 6.12±0.17aA 6.43±0.03aB 6.23±0.10aA 6.61±0.06aA 6.71±0.06aA 5.69±0.09aA 6.36±0.08aA

Means followed by similar lowercase letters in a column are not significantly different from each other (P<0.05); Means followed by similar uppercase letters in a row are not significantly different from each other (P<0.05). EUS: European Standard.

traditional ghee, spiced, untreated and refrigerated time, relatively higher counts of Entero- presented in Table 8. The hedonic rating scale of butter did not significantly (P<0.05) differ from bacteriaceae of 6.70 log cfu/g was observed in color, texture and odor of butter samples each other. While, the mean Entrobacteriaceae spiced butter than in other treatments. This might preserved using traditional ghee; salted, spiced, counts of butter samples for salted, melted and be attributable to poor hygienic status of spices melted, untreated, frozen and refrigerated butter is frozen butter at the end of second months of purchased from open market. Throughout the presented in Figure 1. At the initial time of preservation significantly (P<0.05) differed from preservation period, relatively smaller mean preservation, color, texture and odor of traditional other treatments. However, at the end of third Enterobacteriaceae count was observed in frozen, ghee, spiced, salted, melted, untreated and butter months of preservation, the mean refrigerated butter and traditional ghee compared stored in deep freeze and in refrigerator at 4°C Entrobacteriaceae counts of traditional ghee, to the other treatments. This could be explained in were in acceptable range except for spiced butter spiced, untreated, melted, refrigerated and frozen terms of the inhibitory effects of low storage which was relatively lower compared to others. samples did not significantly differ (P<0.05) except temperatures in refrigerated and frozen butter and This might be attributable to darkening of the color for salted butter. During the initial preservation heat treatment in traditional ghee. Mattick et al. of spiced butter due to mixing with spices such as time, relatively smaller mean Enterobacteriaceae (2001) confirmed that cooling of food to normal black cumin. At initial time of preservation, count of 4.26 log cfu/g of butter was observed in refrigeration temperatures of 0 to 8°C inhibits traditional ghee and salted butter were extremely traditional ghee compared to other treatments. Enterobacteriaceae growth in storage facilitates. liked compared to untreated, butter stored in This might be ascribed due to the heat treatment Rhea (2009) also reported that deep freeze refrigerator at 4°C and in deep freezer which were and moisture removal from butter during ghee retards the growth of undesirable microorganisms liked moderately. The color and odor of traditional making. A report by Mattick et al. (2001) stated and proper salting of butter removes moisture ghee and salted butter were extremely liked. that some thermo tolerant Enterobacteriaceae droplets and negatively affects the growth of Except in spiced butter the color of butter in the comprising a sub-group of mesophiles are undesirable microorganisms. other treatments were extremely liked. capable of growth at up to 44°C, with an optimum growth temperature of 37°C. Fellows (2008) also reported that ghee is preserved by a combination Organoleptic quality of butter preserved using Organoleptic quality of butter at the end of one of heat which destroys enzymes and contaminant traditional preservation techniques month of preservation microorganisms by removing moisture from the butter oil to prevent microorganisms growing Organoleptic quality of butter at the beginning The hedonic rating scale at the end of one month during storage. Samaraweera et al. (2001) of preservation of preservation for color, texture and odor is confirmed that lowering moisture content presented in Table 8. At the end of one month of substantially reduces the growth rate of some The hedonic rating scale at the end of one month preservation, the color, texture and odor of butter Enterobacteriaceae. During the initial preservation of preservation for color, texture and odor is samples for traditional ghee, spiced, salted, 148 Afr. J. Food Sci.

Table 8. Mean scores for descriptive sensory attribute of butter treated under different preservation methods.

Preservation technique Sensory Duration Traditional Spiced Salted Melted Untreated Frozen Refrigerated attribute (months) ghee butter butter butter butter butter butter 0 4.70±0.15a 3.50±0.40b 4.70±0.15a 4.00±0.39ab 4.20±0.25ab 4.20±0.25ab 4.20±0.25ab 1 4.50±0.22a 3.10±0.23b 4.80+0.13a 3.70±0.34b 3.10±0.23b 4.50±0.23a 3.70 ±0.34b Odor 2 4.43±0.20a 2.71±0.36b 3.71±0.36a 2.14±..34bc 1.57±0.30c 4.14±0.26a 3.71 ±0.18a 3 4.85±0.14a 1.71±0.42c 3.43±0.30b 1.71±0.36c 1.43±0.30c 3.86±0.40b 3.57 ±0.20b

0 4.30±0.26a 3.10±0.23b 4.40±0.22a 4.50±0.17a 4.30±0.15a 4.30±0.15a 4.30 ± 0.15 a 1 4.50±0.17a 3.50±0.40b 4.70±0.21a 3.50±0.40b 4.50±0.17a 4.50±0.17a 3.50 ± 0.40 b Texture 2 4.43±0.20a 3.00±0.38bc 3.71±0.42ab 2.71±0.29bc 2.29±0.42c 3.14±0.26b 3.00±0.31b 3 4.71±0.18a 2.57±0.53b 3.43±0.37b 2.43±0.43b 2.29±0.42b 2.86±0.40b 2.86 ±0.34b

0 4.70±0.21a 2.40±0.27c 4.70±0.21a 3.70±0.34b 4.50±0.22a 4.50±0.22a 4.50 ±0.22a 1 4.70±0.15a 2.40±0.27c 4.50±0.22a 3.50±0.40b 3.50±0.40b 4.70±0.21a 4.70 ±0.21a Color 2 4.86±0.14a 1.71±0.36c 4.14±0.14ab 4.14±0.14ab 1.86±0.34c 1.71±0.29c 3.57 ±0.20b 3 4.86±0.14a 1.29±0.18c 3.71±0.47b 1.57±0.30c 1.29±0.18c 3.71±0.52b 3.43 ±0.30b

Means followed by similar letters in a row are not significantly different from each other (P<0.05)

melted, untreated and butter stored in deep freeze and in odor of traditional ghee and salted butter were extreme refrigerator at 4°C were in acceptable range except for and moderate likeness, respectively. While frozen and the color of spiced butter which ranged between dislike refrigerated butter were moderately liked by the sensory slightly to neither like nor dislike. At the end of one panelists. Whereas, the odor and texture of spiced butter month of preservation, the sensory panelists extremely is rated as neither like nor disliked. The color and aroma liked traditional ghee, salted, refrigerated and butter kept of melted and untreated butter were slightly disliked in deep freeze. While, spiced butter was neither liked nor except for their texture. disliked by the sensory panelists. Melted and untreated butter samples were moderately liked. Overall acceptance of traditionally preserved butter at the end of three months Organoleptic quality of butter at the end of two months of preservation The hedonic rating scale on the overall acceptance of color, texture and odor of butter samples at the end of The hedonic rating scale result of color, texture and three months of preservation is presented in Figure 1. aroma of the treatments is presented in Table 8. At the Among all the treatments traditional ghee was extremely end of two months of preservation, the sensory liked followed by salted butter which was rated between acceptability of spiced, melted and untreated butter highly moderate and extreme likeness. At the end of three deteriorated compared to others. Traditional ghee and months, refrigerated and frozen butter were moderately salted butter were rated as extremely liked for their color, liked by the sensory panels. The relative reduction in aroma and texture. This is in agreement with a report of likeness in the overall acceptability of refrigerated and Illingworth et al. (2009) that revealed that application of frozen might be attributable to the change in odor as a heat during preparation of ghee and removal of moisture result of the metabolic and enzymatic activities of and solid non-fat contribute to a product of unique color, psychrophilic bacteria that can multiply under low flavor and texture. temperature.

Organoleptic quality of butter at the end of three CONCLUSION AND RECOMMENDATION months of preservation Major traditional butter preservation methods in the The hedonic rating scale of the butter samples at the end present study are traditional ghee making, spicing, of three months of preservation based on color, texture melting and salting. Major spices used in traditional ghee and odor is presented in Table 8. The color, texture and making are Trachyspermum ammi, Aframomum angusti- Tola et al. 149

Figure 1. Overall acceptability of traditionally preserved butter at the end of three months of preservation.

folium, Nigella sativa, Trigonella foenum, Zingiber Conference of ESAP, Addis Ababa, Ethiopia, 24-26.Pp. 90-104. officinale, Allium sativum and Curcuma domestica. For Eyassu S (2014). Small-scale Milk Processing, Utilization and Marketing of Traditional Dairy Products in North Western Ethiopia. Journal of spicing butter rigonella foenum and Nigella sativa are Food Technology Research, 1(3):122-132. mainly used. Microbial quality of the butter samples were Food and Agriculture Organization (FAO) (1990). The Technology of substandard starting from initial preservation time. Traditional Milk Products in Developing Countries. Animal Production Moreover microbial and organoleptic properties of the and Health Paper. No .85. Rome, Italy, Food And Agriculture Organization Of The United Nations, P 33. samples deteriorated as the storage time elapsed except Food and Agriculture Organization (FAO) (1997). Microbiological for traditional ghee and salting. Hygiene of spices, herbs Analysis. Manual of Food Quality Control. FAO Food and Nutrition and salt used to preserve butter should be kept to reduce Paper, 14/4 Rev. 1. pp. 13-21. contamination. Appropriate application level of spices and Fellows P (2008). Butter and Ghee. In: Rugby, Warwickshire (eds). Technical brief, Practical Action. The Schumacher Center plants materials used per unit weight of butter need to be Technology Development, pp.9-13. optimized. Moreover, comprehensive evaluation including Food safety and standards Authority of India (FSSAI). 2012. Methods of oxidative deterioration of traditionally preserved butter is analysis of foods. vital. Getachew F (2003). A Review of the Small Scale Dairy Sector in Ethiopia. FAO Prevention of Food Losses Programme: Post-harvest

Losses and Food Safety in Sub- Saharan Africa and the Near East.

pp. 26-53. CONFLICT OF INTERESTS Illingworth D, Patil G, Tamime A (2009). Anhydrous Milk Fat Manufacture and Fractionation. In: Tamime, A. Dairy Fats and The authors have not declared any conflict of interests. Related Products. Chichester: John Wiley & Sons Ltd. pp. 108-157 ILSI (International Life Science Institute) (2011). The Enterobacteriaceae and their Significance to the Food Industry. Chris Baylis, Mieke Uyttendaele, Han Joosten and Andy Davies (eds). ILSI ACKNOWLEDGEMENT Europe Report Series. P 52. Joe M, Jayachitra J, Vijayapriya M (2009). Antimicrobial activity of some The authors would like to acknowledge the unreserved common Spices against certain human pathogens. Journal of Medicinal Plants Research, 3(11):1134-1136. efforts made by Holeta dairy laboratory research team Kassahun M (2008). Characterization of milk products consumption during the microbiological analysis and sensory patterns, preference and compositional quality of milk in Ada’a and evaluation. Lume Woreda of East Shoa zone, Central Ethiopia. MSc. Thesis, Hawassa University, Ethiopia. Kilcast D, Subramaniam P (2000). The stability and shelf-life of food. Wood head Publishing Limited, Cambridge. pp. 23-42. REFERENCES Lemma F, Fekadu B, Hegde PB (2004). Rural smallholder milk and dairy products production, utilization and marketing systems in East Ambo District Agricultural Development Office (ADADO). (2014). Showa, Oromia. In: Participatory innovation and research: Lesson for Unpublished quarterly report. livestock development. Proceedings of 12th Annual Conference of Ayantu M (2006). Women’s role on production, processing and ESAP, August 12-14, Addis Ababa, Ethiopia, pp. 17-28. marketing of milk and milk products in Delbo watershed of Wolayta. Livestock Market Development (LMP) (2007). The Livestock Master M.Sc Thesis, Hawassa University, Ethiopia. Plan Study Report. Addis Ababa. Azage T, Tsehay R, Alemu G, Hizkias, K (2000). Milk recording and Mattick, K., Jorgensen, F. and Wang, P. 2001. Effect of challenge herd registration in Ethiopia. Proceedings of the 8th Annual temperature and solute type on heat tolerance of Salmonella 150 Afr. J. Food Sci.

serovars at low water activity. Applied and Environmental Statistical Analysis System (SAS) (2009). Users Guide. Version.9.1. Microbiology, 67:4128-4136. Statistical Analysis System (SAS) Institute, Inc. Carry, NC. Mekdes A (2008). Assessment of processing techniques and quality Seriler H (2003). Yeasts in milk and dairy products. Encyclopedia of attributes of butter produced in Delbo water shade of Wolayita, dairy science. Elsevier science, 4:2761-2769. Southern Ethiopia. M.Sc Thesis. Hawasa University, Ethiopia. P 113. Statistical Packages for Social Sciences (SPSS) (2011). Statistical Michael H, Joseph W, Frank F (2004). Standard Methods for the Packages for Social Sciences. Version. 16. SPSS Corporation. Examination of Dairy Products. 17th ed, American Public Health Tamirat (2007). Physical and Socio Economic Profile of West Shoa Association, Washington DC. USA. pp. 112-118. Zone. BFED Regional data and information core process, Addis Muriuki HG, Thorpe W (2001). Smallholder Dairy Production and Ababa, January 2007. pp. 2-8. Marketing in Eastern and Southern Africa: Regional synthesis. Tsehay R (2001). Milk processing and marketing options for rural small- Nairobi, Kenya. In: Rangnekar D, Thorpe W (2001). Smallholder dairy scale producers. Proceedings of the 5th Annual conference of ESAP. production and marketing-Opportunities and constraints. Proceedings Addis Ababa, Ethiopia. of a South-South Workshop held at NDDB, Anand, India, 13-16 Workneh A, Ulfina G (2011). Gender Role in Peri Urban Dairy O’Mahony F, Peters J (1987). Options for Smallholder Milk Processing Production System of Ambo Town. Journal of Agriculture Extension in Sub-Saharan Africa. International Livestock Center for Africa, and Rural Development, 3(13):224-228. Bulletin 27. Addis Ababa, Ethiopia, pp. 206-247. Yonad (2009). Value chain Analysis of milk and milk products in Borana Quality and Standards Authority of Ethiopia (QSAE). (2009). Ethiopian pastoralist area. unpublished manuscript. Standard. In: Agriculture and Food Technology Technical Committee (eds). Addis Ababa, Ethiopia. P 4. Rhea (2009). Microbial spoilage of dairy products. .In: Standard Methods for examination of dairy products. American Public Health Association, Washington DC. pp. 16-24. Samaraweera I, Buschette L, Rheault D (2001). Survival of pathogenic bacterial organisms in challenge studies of fine granulated sugar, 126:885-889.

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